Vous êtes sur la page 1sur 24

Arch Toxicol (2016) 90:18171840

DOI 10.1007/s00204-016-1744-5

REVIEW ARTICLE

The antioxidant, immunomodulatory, andantiinflammatory


activities ofSpirulina: an overview
QinghuaWu1,2,3 LianLiu4 AncaMiron6 BlankaKlmov3 DanWan7
KamilKuca3,5

Received: 3 January 2016 / Accepted: 24 May 2016 / Published online: 3 June 2016
Springer-Verlag Berlin Heidelberg 2016

Abstract Spirulina is a species of filamentous cyanobacteria that has long been used as a food supplement. In particular, Spirulina platensis and Spirulina maxima are the
most important. Thanks to a high protein and vitamin content, Spirulina is used as a nutraceutical food supplement,
although its other potential health benefits have attracted
much attention. Oxidative stress and dysfunctional immunity cause many diseases in humans, including atherosclerosis, cardiac hypertrophy, heart failure, and hypertension.
Thus, the antioxidant, immunomodulatory, and anti-inflammatory activities of these microalgae may play an important role in human health. Here, we discuss the antioxidant,
* Qinghua Wu
wqh212@hotmail.com
* Kamil Kuca
kamil.kuca@fnhk.cz
1

College ofLife Science, Yangtze University,


Jingzhou434025, Hubei, China

Institute ofBiomedicine, Yangtze University,


Jingzhou434025, Hubei, China

Center forBasic andApplied Research, Faculty


ofInformatics andManagement, University ofHradec
Kralove, Hradec Kralove, Czech Republic

Medical School, Yangtze University, Jingzhou434025,


Hubei, China

Biomedical Research Center, University Hospital Hradec


Kralove, Hradec Kralove, Czech Republic

Department ofPharmacognosy, Faculty ofPharmacy,


University ofMedicine andPharmacy Grigore T. Popa,
Iasi, Romania

Hunan Engineering andResearch Center ofAnimal


andPoultry Science, Key Laboratory ofAgroecological
Processes inSubtropical Region, Institute ofSubtropical
Agriculture, Chinese Academy ofSciences, Changsha, China

immunomodulatory, and anti-inflammatory activities of


Spirulina in both animals and humans, along with the
underlying mechanisms. In addition, its commercial and
regulatory status in different countries is discussed as well.
Spirulina activates cellular antioxidant enzymes, inhibits
lipid peroxidation and DNA damage, scavenges free radicals, and increases the activity of superoxide dismutase
and catalase. Notably, there appears to be a threshold level
above which Spirulina will taper off the antioxidant activity. Clinical trials show that Spirulina prevents skeletal
muscle damage under conditions of exercise-induced oxidative stress and can stimulate the production of antibodies
and up- or downregulate the expression of cytokine-encoding genes to induce immunomodulatory and anti-inflammatory responses. The molecular mechanism(s) by which
Spirulina induces these activities is unclear, but phycocyanin and -carotene are important molecules. Moreover,
Spirulina effectively regulates the ERK1/2, JNK, p38, and
IB pathways. This review provides new insight into the
potential therapeutic applications of Spirulina and may provide new ideas for future studies.
Keywords Spirulina Phycocyanin Antioxidant
Immunomodulation Anti-inflammatory Mechanism
of action
Abbreviations
FDA Food and drug administration
GRAS Generally recognized as safe
COPD Chronic obstructive pulmonary disease
IL Interleukin
TNF Tumor necrosis factor
MAPK Mitogen-activated protein kinase
ROS Reactive oxygen species
MDA Malondialdehyde

13


1818

SOD Superoxide dismutase


CAT Catalase
GPx Glutathione peroxidase
GR Glutathione reductase
PX Peroxidase
APx Ascorbate peroxidase
GSH Glutathione
GST Glutathione S-transferase
DPPH 2,2-Diphenyl-1-picrylhydrazyl
HSC Hepatic stellate cell
HCD High-cholesterol diet
PHGPx Phospholipid hydroperoxide glutathione
peroxidase
6-OHDA 6-Hydroxydopamine
DLM Deltamethrin
NASH Nonalcoholic steatohepatitis
ALT Alanine aminotransferase
OSF Oral submucous fibrosis
COPD Chronic obstructive pulmonary disease
AST Aspartate transaminase
NO Nitric oxide
Se Selenium
Te Tellurium
IFN Interferon
PHA Phytohemagglutinin
LPS Lipopolysaccharide
iNOS Inducible nitric oxide synthase
COX-2 Cyclooxygenase-2
eNOS Endothelial nitric oxide synthase
MPO Myeloperoxidase
NASH Nonalcoholic steatohepatitis
PD Parkinsons disease
AAV9 Adeno-associated virus vector
NMDA 
N-methyl d-aspartate receptor
OSF Oral submucous fibrosis

Introduction
Spirulina, a blue-green algae belonging to the family Oscillatoriaceae, has been used since ancient times as a source
of protein and vitamins (Somchit etal. 2014; Rasool and
Sabina 2009). Spirulina grows naturally in alkaline lakes,
but is commercially produced in large outdoor or greenhouse ponds under controlled conditions (Sotiroudis and
Sotiroudis 2013). Spirulina platensis and Spirulina max
ima are the most common and widely used species and
have been extensively studied in the field of medicine and
by the food industry (Beheshtipour etal. 2012; Hoseini
etal. 2013; Banji etal. 2013). Apart from its protein (60
70%, w/w) and vitamin (4%, w/w) content, Spirulina is
also rich in essential amino acids, minerals, essential fatty
acids, and antioxidants (Kulshreshtha etal. 2008; Soheili

13

Arch Toxicol (2016) 90:18171840

and Khosravi-Darani 2011). Toxicological studies of many


Spirulina species have not identified any toxic effects on the
kidney, liver, or reproductive system in response to acute
or chronic doses (Chamorro etal. 1996, 2002; Kulshreshtha etal. 2008). However, further risk assessment should
be conducted and the production process must be carefully
observed to ensure that no toxic agents are present, specifically heavy metals derived from river water. Normally,
when Spirulina is used as a nutritional supplement, it is
recommended that no more than 15g per day is consumed
(Small 2011).
In recent years, Spirulina has attracted more and more
attention as a potential source of pharmaceutical compounds. Many research studies show that Spirulina has
numerous health benefits, including antioxidant, immunomodulatory, anti-inflammatory, anticancer, anti-viral, and
anti-bacterial activities, as well as positive effects against
hyperlipidemia, malnutrition, obesity, diabetes, heavy
metal chemical-induced toxicity, and anemia (Lee etal.
1998; Lorenz 1999; Hoseini etal. 2013; Kulshreshtha etal.
2008). At present, many studies are focusing on the potential antioxidant activity of Spirulina. Spirulina strongly
induces antioxidant enzyme activity, helps to prevent lipid
peroxidation and DNA damage, and scavenges free radicals
(Abdelkhalek etal. 2015). Moreover, Spirulina protects
against neurotoxicity, hepatonephrotoxicity, and colitis in
animals by reducing oxidative stress (Abdel-Daim etal.
2013, 2015). Thanks to its high antioxidant activity, these
microalgae are considered a promising agent for combating
cardiovascular diseases (Deng and Chow 2010). Although
fewer clinical studies have been performed in humans,
Spirulina does show effective antioxidant activity, making it a potential treatment for chronic obstructive pulmonary disease (COPD) and skeletal muscle damage caused
by exercise-induced oxidative stress (Kalafati etal. 2010;
Ismail etal. 2015). Spirulina also exerts a variety of immunomodulatory and anti-inflammatory activities by regulating key cytokines, including interleukin (IL)-1, IL-2, IL-4,
IL-6, IL-10, and tumor necrosis factor (TNF)- (Mao etal.
2000; El Sheikh etal. 2014; Shokri etal. 2014; Ali etal.
2015).
At present, our understanding of the molecular mechanisms underlying these beneficial effects is limited. Studies
in cell (INS-1E pancreatic beta cells and BV-2 microglial
cells) and rat models show that Spirulina contains phycocyanin and -carotene, both of which are important active
compounds that contribute to its beneficial effects (Schafer
etal. 2002; Li etal. 2009; Chen etal. 2012). The major
ingredients of Spirulina and their related biological properties and specific effects are presented in Table1. Moreover,
mitogen-activated protein kinases (MAPKs) have attracted
much attention as potential mediators of important biological effects. Indeed, Spirulina regulates the ERK1/2,

Hoseini etal. (2013), El Baz etal. (2013) and Berg


etal. (2002)
Inhibition of DNA polymerase activity, decrease in phosphorus demand,
inhibition of production of superoxide anion production
Sulfolipids

Anti-viral, anticancer, anti-inflammatory, anti-proliferative

Youn etal. (2014) and Das (2007)

Antioxidant, anticoagulant,
anticancer, anti-inflammatory,
immunomodulatory, anti-viral,
antinociceptive, anti-hyperlipidemic, anti-hepatotoxic
Antibacterial, antioxidant, antiinflammatory, anticancer, antifibrotic, anti-angiogenic
Sulfated polysaccharide

-Linolenic
acid (GLA)

Antioxidant, anticancer, immunomodulatory, anti-inflammatory

Inhibition of -secretase (BACE1), decrease in antioxidant content of tumor


cells and expression of oncogenes Ras, and Bcl-2, induction of tumor cell
apoptosis by triggering free radical generation, lipid peroxidation and caspase-3 activation

Lorenz (1999), Sudharsan etal. (2015), Ben Abdallah


Kolsi etal. (2015), Hoang etal. (2015) and Wang
etal. (2014a, b)

Shalaby and Shanab (2013), Mallikarjun Gouda etal.


(2015), Bai etal. (2005) and Teng etal. (2016)

1819

-Carotene

Xia etal. (2016), Hwang etal. (2013) and Li etal.


(2009)

Decrease in peroxyl radicals, lipid peroxidation, total cholesterol (CHOL),


low-density lipoproteins (LDL), COX-2, iNOS, ROS, MDA, TNF-, IL-6,
and IL-1
Inhibition of NF-B nuclear translocation, protection against singlet oxygenmediated lipid peroxidation, and intracellular accumulation of ROS, inhibition of expression of inflammatory genes
DNA damage repair, protection against hypoxia injury, decrease in oxidative
stress (superoxide and hydroxyl radicals scavenging effects, inhibition of
lipid peroxidation), iNOS, TNF-, IL-6, IL-8, plasma triacylglycerol (TG),
total cholesterol (TC), plasma low-density lipoprotein (LDL) levels
Antioxidant, anticancer,
anti-inflammatory
Phycocyanin

References
Specific effects
Biological properties
Ingredient

Table1Summary of the biological properties and specific effects of the major ingredients of Spirulina

Arch Toxicol (2016) 90:18171840

JNK, p38, and IB signaling pathways to exert its antioxidant, immunomodulatory, and anti-inflammatory activities
(Khan etal. 2006; Yogianti etal. 2014).
Several excellent reviews provide valuable knowledge
about the nutritional value and medical uses of Spirulina
(Kulshreshtha etal. 2008; Small 2011; Deng and Chow
2010; Hoseini etal. 2013; Sotiroudis and Sotiroudis 2013).
The nutritional value of Spirulina is well described by
Small (2011); however, the review does not mention medical applications. The antioxidant, immunomodulatory, and
anti-inflammatory activities of the microalgae are only
summarized briefly by Kulshreshtha etal. (2008), Hoseini
etal. (2013), and Sotiroudis and Sotiroudis (2013), who
unfortunately do not provide a global overview of these
activities. The hypolipidemic, antioxidant, and anti-inflammatory activities are well summarized by Deng and Chow
(2010). However, during the last 5years, we have learned
much about the antioxidant, immunomodulatory, and antiinflammatory activities of Spirulina. Moreover, the mechanisms underlying these activities are rarely addressed in
earlier reviews.
Thus, the aim of this review article is to update and
critically review data related to the antioxidant activities of
Spirulina. These data were obtained from both invitro and
invivo studies. We will summarize the immunomodulatory and anti-inflammatory effects of Spirulina. Moreover,
we will examine the mechanisms underlying the antioxidant, anti-inflammatory, and immunomodulatory activities.
Finally, the commercial and regulatory status of Spirulina
in different countries is discussed. We hope this review
will further our understanding of the therapeutic potential
of Spirulina in terms of human disease and clarify data
obtained from functional food studies.

Methods
This review was written after searching several databases,
including Web of Science, Elsevier Science Direct, ProQuest, Springer, Scopus, and Emerald, for papers published
between 1999 and 2016. The searches were performed
using the following key words/phrases: Spirulina, phycocyanin, phycocyanobilin, Spirulina and -carotene, Spirulina
and antioxidant, Spirulina and oxidative stress, Spirulina
and immunomodulation, Spirulina and anti-inflammatory,
and Spirulina and mechanism of activity, Spirulina and
commercial, Spirulina and regulatory. The relevant publications were selected as follows:
1. Identification: identification of key words and, consequently, the availability of the relevant sources;
2. Checks to eliminate duplication;

13


1820

3. Assessment of relevancy: verification (on the basis of


reading abstracts) that the selected study corresponded
to the set goal;
4. Assessment of the selected studies.
Duplication of articles was mainly encountered when
using the Web of Science and Scopus databases. After close
examination of the abstracts, articles that contained the key
words but did not address the set goal were excluded. Studies outside the selected time frame and those that reported
the same findings or reported findings that were outdated
were also excluded.

Results anddiscussion
Antioxidant activity
Reactive oxygen species (ROS) attack and damage molecules in biological systems, leading to various disorders
and diseases. Thus, oxidative stress plays a role in many
diseases, including hypertension, diabetes mellitus, atherosclerosis, ischemic disease, and malignancy. The lipid
peroxides (LPOs) malondialdehyde (MDA) and 4-hydroxynonenal are important markers of oxidative stress (Yoshikawa and Naito 2002). Recently, much attention has been
paid to the antioxidant potential of Spirulina species. Many
invitro and invivo studies show that treatment with Spirulina significantly reduces oxidative stress. These antioxidant and protective effects are mediated by phycocyanins,
-carotene, and other vitamins and minerals contained
within Spirulina (Abdel-Daim etal. 2013; Upasani and
Balaraman 2003).
In vitro studies
A number of invitro studies have identified the potential
antioxidant activity of Spirulina or its extracts. Qing etal.
(2003) found that the activities of two antioxidant enzymes
contained within Spirulina maxima, superoxide dismutase
(SOD) and catalase (CAT), increased under conditions of
light stress, although SOD was more sensitive to external
stress than CAT. Two other studies report a more complicated relationship between the antioxidant activity of
Spirulina and these antioxidant enzymes (Bermejo-Bescs
etal. 2008; Abd El-Baky etal. 2009). Bermejo-Bescs
etal. (2008) examined the ability of Spirulina platensis to
inhibit lipid peroxidation and its effects on the activities of
SOD, CAT, glutathione peroxidase (GPx), and glutathione
reductase (GR) after exposure of SH-SY5Y neuroblastoma
cells to iron-induced oxidative stress. Spirulina platensis
maintained the activity of cellular antioxidant enzymes
(total GPx, GPx-Se, and GR) and increased the levels of

13

Arch Toxicol (2016) 90:18171840

reduced glutathione in these cells. Intriguingly, the antioxidant capacity of Spirulina platensis could be enhanced
when exposed to additional environmental stress (Abd ElBaky etal. 2009). For example, Abd El-Baky etal. examined the possibility of increasing the amounts of some
bioactive compounds in Spirulina platensis by cultivating
cells in medium supplemented with varying concentrations of hydrogen peroxide (H2O2). They found a positive
correlation between increasing H2O2 concentrations and
increasing amounts of cellular lipophilic antioxidants (total
carotenoids and -tocopherol) and hydrophilic antioxidants
(glutathione and ascorbic acid). In addition, increasing
H2O2 levels led to a significant linear increase in the activities of antioxidant enzymes in Spirulina platensis, including CAT, peroxidase (PX), SOD, and ascorbate peroxidase
(APx). Thus, Spirulina platensis may be useful for treating
diseases that are aggravated by ROS and for developing
novel treatments for neurodegenerative disorders such as
Alzheimers or Parkinsons disease.
Liver fibrosis is a chronic disease, the therapeutic strategy for which involves inhibiting the proliferation of activated hepatic stellate cells (HSCs) and the subsequent
induction of HSC apoptosis (Wu etal. 2005). Based on this
idea, Wu etal. (2005) examined whether the antioxidant
effects of Spirulina inhibited proliferation of HSCs. Spirulina did inhibit proliferation of HSCs and induced apoptosis after 12h. Similar results were reported by Konckov
etal. (2014) who used invitro cell-based models to examine the anticancer effects of Spirulina platensis. The results
showed that Spirulina not only had anti-proliferative
effects, but also inhibited the production of mitochondrial
ROS and affected glutathione redox status.
Iron accumulation in the brain is involved in the pathogenesis of neurodegenerative diseases, most likely through
iron-induced oxidative damage to neuronal cells (Sagara
etal. 2015). Recently, Japanese scientists (Sagara etal.
2015) reported that Spirulina protects PC12 cells against
iron-induced toxicity. They also found that extracts of
Spirulina showed antioxidant and free radical-scavenging
activities in these cells. Thus, Spirulina extracts may protect against neurodegenerative disorders caused by excessive iron accumulation in the brain.
In addition to cell lines, studies have used tissue
homogenates to assess the antioxidant activity of Spirulina.
Miranda etal. (1998) measured the antioxidant activity in a
brain homogenate incubated with Spirulina or with thiobarbituric acid reactant substances (TBARS; used as a control)
for 1h at 37C. Peroxidation of the rat brain homogenate
was inhibited by almost 95% in the presence of 0.5mg of
the Spirulina-containing methanolic extract.
Taken together, the above studies suggest that Spirulina
has potent antioxidant activity invitro and that this activity
is due to its ability to activate cellular antioxidant enzymes

Arch Toxicol (2016) 90:18171840

such as GPx, GPx-Se, and GR. Furthermore, its antioxidant


activity is strongly correlated with the activity of scavenging enzymes such as SOD, CAT, PX, and APx, as well as
other antioxidants such as GSH and -carotene. Notably,
the antioxidant activity is enhanced by exposure to additional environmental stress. These early invitro studies
provided important results that guided the design of subsequent invivo studies in animals and humans.
Animal studies
A number of animal-based studies have examined the
antioxidant activity of Spirulina. Kim etal. (2010) used a
hypercholesterolemic New Zealand white rabbit model to
study the beneficial effects of Spirulina platensis against
tissue lipid peroxidation and oxidative DNA damage.
Hypercholesterolemia was induced by feeding rabbits a
high-cholesterol diet (HCD) for 4weeks. The diet was then
supplemented with Spirulina (1 or 5%, w/w) for 8weeks.
The authors found that increased lipid peroxidation
induced by a HCD was significantly attenuated by Spirulina supplementation. Indeed, lipid peroxidation returned
to control levels. Supplementation with 1 or 5% Spirulina
induced GSH, GPx, and GR activity in the liver. In addition, the level of DNA damage in lymphocytes was significantly reduced. This suggests that dietary supplementation
with Spirulina is a promising strategy to protect the cells
from lipid peroxidation and DNA damage caused by oxidative stress. Another study examined the antioxidant and
therapeutic effects of Spirulina in a Balb/c mouse model
of trichloroethylene-induced cutaneous irritation (Gurbuz
etal. 2013). Moreover, combining Spirulina with other
products such as whey protein increased protection against
the CCl4-induced oxidative stress response in rats (Gad
etal. 2011; Hassan etal. 2012).
The major mechanism responsible for mycotoxininduced toxicity is induction of oxidative stress (Wu etal.
2014a, b; Dohnal etal. 2014). Hassan etal. (2012) evaluated the ability of Spirulina to regulate gene expression and
counteract oxidative stress in rats with mycotoxin-induced
aflatoxicosis. Supplementation with Spirulina led to a significant reduction in aflatoxin-induced oxidative stress,
as revealed by reduced lipid peroxidation, increased GSH
content, and upregulated expression of the gene encoding phospholipid hydroperoxide glutathione peroxidase
(PHGPx). Spirulina also inhibited DNA damage, as indicated by downregulation of Fas gene expression, reduced
DNA fragmentation, and reduced numbers of micronucleated erythrocytes.
The antioxidant activity of Spirulina protects animals
against neurotoxicity. Tobn-Velasco etal. (2013) studied
the effects of Spirulina maxima against 6-hydroxydopamine, which is neurotoxic to the rat striatum. Rats received

1821

6-OHDA (700mg/kg/day b.w.) for 20days via a single


injection into the dorsal striatum. The striatum was collected, and the levels of nitric oxide (NO) and ROS, lipid
peroxidation, and mitochondrial activity were measured:
All increased significantly after exposure to 6-OHDA. Rats
also showed reduced locomotor activity. However, rats
treated with Spirulina maxima showed restored locomotor
activity and reduced levels of NO, ROS, and LPO.
Humans and animals exposed to deltamethrin (DLM)
can develop hepatonephrotoxicity. Moreover, rats treated
with DLM show a significant increase in renal lipid peroxidation and reduced expression of antioxidant biomarkers
(Abdel-Daim etal. 2013). Supplementation with Spirulina
inhibited lipid peroxidation. In addition (and as reported
by previous invitro studies), Spirulina induced a significant dose-dependent increase in the activity of the antioxidant enzymes in the kidneys and liver, as well as increased
glutathione levels. It is possible that Spirulina protects
against DLM-induced oxidative stress either directly (by
inhibiting lipid peroxidation and scavenging free radicals) or indirectly (by increasing the activity of SOD and
CAT) (Abdel-Daim etal. 2013). Subsequently, AbdelDaim etal. (2015) showed that Spirulina had a significant
modulatory effect on acetic acid-induced colitis in rats by
increasing antioxidant enzyme activity, reducing lipid peroxidation, and reducing expression of inflammatory markers. Recently, Ibrahim and Abdel-Daim (2015) reported
that Spirulina minimizes the toxic effects of tilmicosin in
mice by scavenging free radicals and exerting potent antioxidant activities. Mice showed a significant increase in
serum cardiac injury biomarkers and cardiac lipid peroxidation when treated with tilmicosin for 5days. Moreover,
evidence suggested that expression of antioxidant biomarkers was inhibited. However, treatment with Spirulina plat
ensis (1000mg/kg) for 5days normalized the increased
serum levels of lactate dehydrogenase and creatine kinase
(CK). Furthermore, Spirulina platensis reduced tilmicosininduced lipid peroxidation and oxidative stress in a dosedependent manner.
The presence of silicon-enriched Spirulina in the diet
may protect the heart and liver against oxidative stress by
reducing the activity of nicotinamide adenine dinucleotide
phosphate oxidase and by maintaining SOD and GPx activity (Vid etal. 2015a). Moreover, dietary supplementation with silicon-enriched Spirulina had no harmful effects
in rats. Thus, dietary silicon-enriched Spirulina may be a
useful treatment for conditions caused by oxidative stress
(Vid etal. 2015b).
Oxidative stress and the relationship between inflammation and immunity play a critical role in the pathogenesis
of nonalcoholic steatohepatitis (NASH) (Sanyal 2001; Pak
etal. 2012). Rat models of NASH show normalization of
an imbalance between CD4+ and CD8+ cells, increased

13


1822

production of ROS by liver mitochondria and leukocytes,


and activation of NF-B (Pak etal. 2012). Administration
of Spirulina led to significant abrogation of these phenomena. This may be because Spirulina dampens the inflammatory response through antioxidative and anti-inflammatory
mechanisms and by breaking the crosstalk between signaling pathways related to oxidative stress and inflammation
(Pak etal. 2012).
Currently, Spirulina has attracted attention due to its
growth-promoting effects and antioxidant potential in fish
(Abdelkhalek etal. 2015). Administration of Spirulina to
freshwater fish Nile tilapia minimized the toxic effects of
DLM via its free radical-scavenging and antioxidant activities. A study by Abdelkhalek etal. (2015) showed that
DLM reduced the levels of GSH and GSH-Px in fish tissue and reduced the activity of SOD and CAT. However,
Spirulina supplementation significantly increased the levels of all of these antioxidant biomarkers. Fish and squid
surimi have many health benefits (Kris-Etherton etal.
2003). Indeed, glucomannanSpirulina-enriched squid
surimi increases SOD, CAT, and GPx levels in Fa/Fa rats
(Vzquez-Velasco etal. 2014). Spirulina is one of the most
widely used microalgae species in aquafeeds and as such
shows high dietary antioxidant activity in aquatic species
such as the juvenile olive flounder (Kim etal. 2013; Nakagawa and Montgomery 2007).
Muga and Chao (2014) used hypercholesterolemic hamsters to examine the effects of Spirulina on oxidative stress.
They noticed that high cholesterol levels increased oxidative stress and inhibited the production of antioxidants.
Treatment of hamsters with Spirulina inhibited lipid peroxidation and restored antioxidant status. Moreover, Spirulina
induced significant increases in hepatic GSH levels, and
in GSHPx and SOD activity. Spirulina platensis not only
reduces damage caused by oxidative stress and reduces
accumulation of amyloid -protein, but also increases
antioxidant activity in senescence-accelerated mice, all of
which might play an important role in preventing memory
loss, suggesting a possible beneficial role in human Alzheimers disease (Hwang etal. 2011a, b).
Notably, there seems to be a threshold concentration
above which the antioxidant activity of Spirulina tapers
off. Ding etal. (2004) found that treating liver-injured mice
with Spirulina for 45days led to a reduction in alanine
aminotransferase (ALT) activity and MDA content, while
at the same time increasing SOD activity. However, preliminary observations suggest that exceeding a certain dose
leads to a reduction in antioxidant activity; further studies
should examine the threshold concentrations of Spirulina to
determine whether this is really the case.
In summary, several animal studies show that Spirulina
has potent antioxidant activity. Similar to observations
made in invitro studies, animal studies show that Spirulina

13

Arch Toxicol (2016) 90:18171840

increases the activity of antioxidant enzymes, reduces lipid


peroxidation and DNA damage, and scavenges free radicals. Moreover, when combined with other products, Spirulina is even more effective at reducing oxidative stress. In
addition, Spirulina protects against neurotoxicity, hepatonephrotoxicity, and colitis. Recent studies show that Spirulina could be an effective supplement in fish feed due to
its promising antioxidant potential. It is possible that there
is a threshold value above which Spirulina shows reduced
antioxidant activity, but further studies are needed to verify
this. Finally, Spirulina supplementation may be useful for
reducing oxidative stress induced by mycotoxins; therefore,
it may be a promising treatment for mycotoxin-induced
intoxication in both animals and humans.
Effects onheavy metalinduced oxidative stress
Exposure to heavy metals (such as mercury, lead, and
chromium) induces ROS and an associated oxidative
stress response (Farina etal. 2004; Sharma etal. 2007,
2013; Elshazly etal. 2015). Numerous invivo studies
have explored the potential of Spirulina to combat oxidative stress induced by heavy metals. Spirulina fusiformis
protected Swiss albino mice against mercuric chlorideinduced oxidative stress (Sharma etal. 2007). Oxidative
stress induced by mercuric chloride increased lipid peroxidation and subsequently reduced the amount of GSH and
antioxidant enzymes in the liver. Supplementation with
Spirulina (800mg/kg b.w.) for 40days reduced lipid peroxidation. Moreover, the activities of SOD, CAT, and GST
were restored to near normal levels. In addition to mercury,
lead may also cause oxidative damage. Dar etal. (2014)
studied the antioxidant effects of Spirulina by supplementing the diet of catfish with Spirulina platensis. They found
that Spirulina had beneficial effects against cadmium chloride (CdCl2)-induced nephrotoxicity. Spirulina platensis is
a potent antioxidant that ameliorates chromium-induced
nephrotoxicity in rats and provides protection by increasing
antioxidant activity and reducing oxidative stress (Elshazly
etal. 2015).
An increasing number of studies have focused on the
effects of Spirulina against lead-induced oxidative stress
in animals and humans. Jeyaprakash and Chinnaswamy
(2007) evaluated the antioxidant properties of Spirulina in
albino rats. When rats were treated with lead acetate for
30days, their levels of GSH, CAT, SOD, and GPx were
much lower than those in control rats. Administration of
Spirulina resulted in a marked recovery of these parameters, suggesting that its antioxidant effects protect against
lead-induced toxicity. Similarly, El-Tantawy (2015) examined the antioxidant activity of a Spirulina supplement in
rats with lead acetate-induced hepatic injury. Lead intoxication resulted in a significant increase in serum aspartate

Arch Toxicol (2016) 90:18171840

transaminase (AST), caspase-3, MDA, and NO levels and


ALT activity, and a pronounced decline in GSH and SOD
activity. Spirulina supplementation resulted in significant
antioxidant and anti-apoptotic activity. Ponce-Canchihuamn etal. (2010) also reported that Spirulina protected rat
liver and kidneys against lead-induced oxidative stress.
Thus, animal studies suggest that Spirulina is a potential
antidote for heavy metal toxicity, particularly that induced
by lead. However, no clinical studies designed to examine
the efficacy of Spirulina against heavy metal toxicity have
been performed.
Critical roles ofphycocyanin/phycocyanobilin in
the antioxidant activity
Spirulina platensis is rich in phycocyanin. The deep
blue color of phycocyanin is due to the open tetrapyrrole
chromophore, phycocyanobilin, which is covalently linked
to the apoprotein (Fig.1) (Hirata etal. 1999, 2000). Allophycocyanin is also an important phycobiliprotein in Spir
ulina platensis because it transfers energy to chlorophyll
molecules (Cherdkiatikul and Suwanwong 2014). The
promising antioxidant and free radical-scavenging properties of Spirulina platensis may be due to its phycocyanin
content, which in turn attributes its antioxidant activity to
phycocyanobilin (Hirata etal. 2000; Sagara etal. 2015).
Hirata etal. (2000) examined the ability of phycocyanobilin isolated from Spirulina platensis to protect methyl
linoleate and phosphatidylcholine liposomes from oxidation. Phycocyanobilin effectively inhibited the peroxidation
of methyl linoleate and the oxidation of phosphatidylcholine
liposomes. It is noteworthy that the antioxidant activity of
phycocyanobilin was almost the same as that of phycocyanin when the concentrations of the two molecules in the
reaction mixture were adjusted to yield equivalent phycocyanobilin concentrations. Thus, phycocyanobilin appears to
be responsible for most of the antioxidant activity attributed
to phycocyanin. Another study by Bermejo etal. (2008)
reported that phycocyanin appears to inhibit the generation of hydroxyl and peroxyl radicals, as well as lipid peroxidation. Moreover, when selenium (Se) or tellurium (Te)
bind to phycocyanin, its antioxidant activity is significantly
increased (Huang etal. 2007; Yang etal. 2014). A rat model
of nonalcoholic steatohepatitis revealed that phycocyanin
administration significantly reduced the production of ROS
in liver mitochondria and leukocytes (Pak etal. 2012).
Zhou etal. (2005) examined how different factors affect
the antioxidant activity of phycocyanins. While phycocyanins generated hydroxyl radicals in the light, they scavenged them in the dark. Also, when phycocyanins were
denatured, they lost their ability to generate hydroxyl radicals but became much more efficient at scavenging them.
Furthermore, trypsin-catalyzed hydrolysis of phycocyanins

1823

revealed that the apoprotein part of the molecule made the


greatest contribution to its antioxidant activity.
Both phycocyanin and phycocyanobilin from Spirulina
platensis protect against diabetic nephropathy by inhibiting oxidative stress (Zheng etal. 2013). Oral administration of phycocyanin (300mg/kg b.w.) for 10weeks normalized urinary markers of renal oxidative stress and the
expression of NADPH oxidase in db/db mice. Similar
antioxidant effects were also observed following administration of phycocyanobilin (15mg/kg b.w.) for 2weeks. In
addition to phycocyanin, allophycocyanin possesses strong
antioxidant activity. Further studies showed that allophycocyanin is superior to phycocyanin in terms of scavenging
peroxyl radicals, whereas phycocyanin is better at scavenging hydroxyl radicals (Cherdkiatikul and Suwanwong
2014). Other studies show that treatment with phycocyanin leads to a marked reduction in expression of oxidative
stress and apoptotic markers, astroglial activation, and cell
death caused by tributyltin chloride (TBTC) (Mitra etal.
2015). Thus, phycocyanin is an important active compound
involved in Spirulina-mediated antioxidant activity. The
antioxidant effects of Spirulina in both animals and humans
are summarized in Table2.
Immunomodulatory activity
Animal studies show that Spirulina is a powerful stimulator
of the immune system. It does this by increasing the phagocytic activity of macrophages, causing NK cells to accumulate in tissues, by stimulating the production of antibodies
and cytokines, and by activating and mobilizing T and B
cells (Qureshi and Ali 1996; Mao etal. 2000; Gad etal.
2011; Khan etal. 2005).
Spirulina has marked immunomodulatory effects. Mao
etal. (2000) used human peripheral blood mononuclear
cells (PBMCs) to evaluate the immunomodulatory activity
of Spirulina. They showed that Spirulina increased IL-1,
IL-4, and interferon (IFN)- production by PBMCs by
almost 2.0, 3.3, and 13.6 times basal levels, respectively. El
Sheikh etal. (2014) showed that treatment of PBMCs with
phycocyanin from Spirulina platensis led to a significant
increase in IFN- production, but differently from Mao
etal. (2000) they found that phycocyanin reduced IL-4 production. These differences may be due to the presence of
other active compounds in the Spirulina extracts.
An invivo study showed that when mice suffering from
systemic candidiasis and breast cancer were treated with
Spirulina platensis (800mg/kg b.w.) for 3days, the expression of genes encoding IL-4 and IL-10 was significantly
reduced, whereas that of genes encoding IL-17, TNF-,
and IFN- was markedly increased (Shokri etal. 2014).
The results of this study suggest that Spirulina platensis
might exert tumoricidal effects invivo by increasing the

13


1824

Arch Toxicol (2016) 90:18171840

Fig.1Chemical structure of phycocyanin, phycocyanobilin, and bilirubin

production of pro-inflammatory TNF- and IFN-; however, further investigations are needed to fully evaluate the
efficacy of Spirulina platensis in tumor-bearing mice.
Recently, Chen etal. (2014) purified phycocyanin from
Spirulina platensis cultured using power plant flue gas.
The immunomodulatory activity of the purified molecule
was then examined using the murine macrophage cell line,
J774A. They found that phycocyanin increased the secretion of TNF-, IL-1, and IL-6 by J774A cells, along with
the expression of IL-1 and COX-2 proteins. In addition,
both Spirulina platensis and phycocyanin inhibited expression of inflammation-related genes by lipopolysaccharide
(LPS)-stimulated BV-2 microglial cells. In an earlier study,
Chen etal. (2012) showed that LPS upregulated the expression of genes encoding inducible nitric oxide synthase
(iNOS), cyclooxygenase-2 (COX-2), TNF-, and IL-6 in
BV-2 microglial cells. However, exposure to Spirulina or
phycocyanin led to a significant reduction in the expression of these genes. Similar results were reported by Sin
etal. (2014) and Ali etal. (2015), who found that Spirulina
maxima inhibited expression of genes encoding TNF- and
IL-6 in the human cell line CCD-986sk. However, TNF-
gene expression was significantly upregulated when rats
were fed a diet supplemented with glucomannanSpirulina (Vzquez-Velasco etal. 2014). Spirulina also affects
the immune response in healthy, elderly male and female
human subjects. Spirulina supplementation resulted in a
significant rise in plasma IL-2 concentrations in both male
and female subjects, along with a marked reduction in IL-6
concentrations (Park etal. 2008).

13

Spirulina fusiformis also showed marked immunosuppressive effects in a series of invivo and invitro studies
(Rasool and Sabina 2009). Mice treated with Spirulina
fusiformis (400 or 800mg/kg) showed significant suppression of immune responses, delayed-type hypersensitivity
reactions, and TNF- production. In vitro, Spirulina fusi
formis (50 or 100mg/mL) reduced mitogen-induced proliferation of T lymphocytes in a concentration-dependent
manner. This situation mimics unrestricted proliferation of
T cells invivo, which occurs in diseases such as arthritis.
Thus, Spirulina fusiformis may form the basis of alternative
treatments for arthritis. Another invivo study showed that
mice treated with salicylate showed a significant increase
in expression of mRNA for N-methyl d-aspartate receptor
subunit NR2B, TNF-, and IL-1 mRNA in the cochlea
and colliculus (Hwang etal. 2013). However, dietary supplementation with Spirulina or phycocyanin led to a significant downregulation of the expression of these immuneand inflammatory-related genes.
Anemia and immunosenescence are common in older
subjects. A working group led by Dr. Gershwin (Selmi
etal. 2011) suspected that Spirulina might ameliorate
anemia and immunosenescence in senior citizens. Therefore, they treated volunteers (>50years old) with Spirulina
for 12weeks. They found that the levels of corpuscular
hemoglobin had increased in both sexes after this time.
Moreover, the majority of subjects showed an increased
indoleamine 2,3-dioxygenase enzyme activity, a sign of
immune function, along with an increased white blood cell
count, after 6 and 12weeks of Spirulina supplementation,

H2O2

Human islet amyloid polypeptide

Medium containing
H2O2

INS-1E pancreatic
beta cells

Microsomal fraction

Human pancreatic
cancer cells

Oxygen radical
absorbance capacity
(ORAC) assay
PC12 cells

HepG2 cells

EDTA

Iron

2,2-Azino-bis(3ethylbenzothiazoline-6-sulfonic
acid

Phorbol12-myristate13-acetate

Cell-free and cellbased incubation

In vitro DPPH radicalscavenging system

The peroxidation of methyl


linoleate was inhibited and
oxidation of phosphatidylcholine
liposomes was controlled
Dose-dependent
inactivation of superoxide radical
generated

Methyl linoleate

Hydrophobic system
or phosphatidylcholine liposomes

In vitro

12h
12h

30min

30min

14days

30min

Te-Spirulina platensis 25200g/mL

0.951.81M

550mg/mL

0.21.6mg/mL

1mM

Allophycocyanin and
phycocyanin

24h

24h

60min

200400min

Duration

100200g/mL

Spirulina platensis

2.55M

Phycocyanin

501000g/mL

Zn-Spirulina

28mM

10100M

Phycocyanobilin

Spirulina platensis

Dosing

Component

Spirulina platensis
Prevented the free radical-evoked
DNA degradation and showed
antioxidant and free radical-scavenging activities
Decreased generation of mitochon- Spirulina platensis
drial ROS and glutathione redox
status
The generation of hydroxyl radical, Spirulina platensis
the production of peroxyl radicals, and the lipid peroxidation
process were inhibited

Spirulina platensis behaved with


different strategies against H2O2induced oxidative stress. Their
response strongly correlated with
the scavenging enzymes SOD,
CAT, PX, and APx
The overproduction of intracellular ROS and MDA, as well as
changes in activities of SOD and
GSH-Px enzymes were significantly prevented
Scavenging free
radicals
Apoptotic cell death
and nuclear condensation induced
by tert-butyl hydroperoxide in
HepG2 cells were significantly
attenuated
Scavenging peroxyl, hydroxyl
radicals

Specific effects

Inducing agent

Object

Experimental
model

Table2Antioxidant activity of Spirulina and its active compounds evaluated in invitro systems, animals, and humans

Bermejo etal. (2008)

Konckov etal. (2014)

Sagara etal. (2015)

Cherdkiatikul and
Suwanwong (2014)

Shalaby and Shanab


(2013)
Yang etal. (2014)

Li etal. (2009)

Abd El-Baky etal. (2009)

Dartsch (2008)

Hirata etal. (2000)

References

Arch Toxicol (2016) 90:18171840


1825

13

13
Lead acetate

Mercuric chloride

Lead acetate

Rat

Swiss albino mice

Rat

CCl4

Rat

Deltamethrin

Senescence-accelerated
prone-8 (SAMP8)
mice
Rat
Aflatoxins

Rat

High-cholesterol
New Zealand white
diet
rabbit with hypercholesterolemia

CCl4

Rat

Animals

Inducing agent

Object

Experimental
model

Table2continued

40days

30days

8weeks

800mg/kg b.w

1g/day/rat

Diet with 5% Spirulina

Spirulina platensis

Spirulina maxima

Spirulina fusiformis

30days

10mg/kg b.w

Spirulina

It reduced lipid peroxidation and


Spirulina platensis
oxidative stress in a dose-dependent manner

30days

1week

30days

5days

0.5g/day/rat

50200mg/kg b.w

50mg/kg b.w

1000mg/kg b.w

45days

106385mg/kg b.w

Spirulina

Duration

Dosing

Component

Spirulina
Antioxidant, radical scavenging,
and metal-chelating activities
in dose-dependent manner were
observed
The lipid peroxidation and amyloid Spirulina platensis
b-protein deposition was significantly reduced
Decreased lipid peroxidation level, Spirulina
increased GSH content and
upregulated PHGPx gene expression; inhibited DNA damage, and
decreased the percentage of DNA
fragmentation

Reduced ALT activity, MDA


content, improved SOD activity;
when the dosage of Spirulina
overruns a certain value, the
antioxidant ability of Spirulina
was reduced
Spirulina significantly increased the
levels of GSH, SOD, CAT, and
GPx, which were reduced by lead
acetate
Activities of antioxidant enzymes
SOD, CAT, and GST were
restored to near normal level by
Spirulina supplementation
The significant changes on plasma
and liver lipid levels and on the
antioxidant status of the liver and
kidney were prevented
The increased lipid peroxidation
level were reduced to control
values; oxidative stress biomarkers such as GSH, GPx, GR, and
GST were significantly improved;
DNA damage in lymphocytes
were significantly reduced

Specific effects

Abdel-Daim etal. (2013)

Hassan etal. (2012)

Hwang etal. (2011a, b)

Gad etal. (2011)

Kim etal. (2010)

Ponce-Canchihuamn
etal. (2010)

Sharma etal. (2007)

Jeyaprakash and
Chinnaswamy (2007)

Ding etal. (2004)

References


1826
Arch Toxicol (2016) 90:18171840

Humans

Experimental
model

Trichloroethylene

Flurode

Mice

Mice

Human

Rat

Rat

Mice

Fish Nile tilapia

Hamster

Mice

Rat

6-OHDA

Rat
Levels of MDA and NO were
decreased, whereas SOD activity
was increased

Decreased levels of NO, ROS, and


lipoperoxidation in the striatum

Specific effects

3weeks

15g/day

5days

1000mg/kg b.w

4days

28days

1.46g/L

50mg/kg b.w

4weeks

11mg/kg feed

4weeks

10weeks
2weeks

300mg/kg b.w
15mg/kg b.w

500mg/kg b.w

15days

250 and 500mg/


kg b.w

7days

7days

60mg/day

Spirulina

40days

Duration

230mg/kg b.w

700mg/kg b.w

Dosing

Spirulina maxima

Component

Hematological disorders and oxida- Spirulina


tive stress were reduced
Flurode
Spirulina augmented the displace- Spirulina platensis
ment of fluoride, facilitated antioxidant formation, and protected
Purkinje cells
Phycocyanin phycoDiabetic nephropa- Urinary and renal oxidative stress
cyanobilin
thy
markers and the expression of
NADPH oxidase components
were normalized
High-cholesterol
Reduced lipid peroxidation level to Spirulina
diet
near or below control levels, and
increased SOD activity
Spirulina platensis
Deltamethrin
The reduction of GSH level
induced by deltamethrin was
reversed; GSH-Px and SOD
activity were increased significantly
Tilmicosin
Tilmicosin-induced lipid peroxida- Spirulina platensis
tion and oxidative stress were
reduced in a dose-dependent
manner
Lead acetate
GSH level and SOD activity in liver Spirulina
homogenate were increased
Phycocyanin
Tributyltin chloride Phycocyanin remarkably reduced
oxidative stress and apoptotic
markers, astroglial activation and
cell death
Spirulina platensis
Exercise-induced
Plasma concentrations of MDA
oxidative stress
were significantly decreased;
the activity of blood SOD was
significantly raised

Inducing agent

Object

Table2continued

Lu etal. (2006)

Mitra etal. (2015)

El-Tantawy (2015)

Ibrahim and Abdel-Daim


(2015)

Abdelkhalek etal. (2015)

Muga and Chao (2014)

Zheng etal. (2013)

Banji etal. (2013)

Sharma etal. (2013)

Tobn-Velasco etal.
(2013)
Gurbuz etal. (2013)

References

Arch Toxicol (2016) 90:18171840


1827

13

Ismail etal. (2015)


60days
12g/day
Spirulina

Exercise

Cadmium chloride

Human

Patients with COPD

Serum content of MDA, lipid


hydroperoxide, and cholesterol
were significantly reduced, while
increasing GSH, and the activity
of SOD

Inducing agent
Object

Spirulina
Spirulina decreased carbohydrate
oxidation and increased fat
oxidation rate, attenuated the
exercise-induced increase in lipid
peroxidation

6g/day

4weeks

Kalafati etal. (2010)

Arch Toxicol (2016) 90:18171840

Experimental
model

Table2continued

Specific effects

Component

Dosing

Duration

References


1828

13

respectively. Thus, Spirulina appears to ameliorate anemia


and immunosenescence in older subjects. However, randomized clinical trials are needed to determine whether
Spirulina supplementation is really beneficial. In addition,
Spirulina preparations contain immunoactive insulin; therefore, it may have a potentially beneficial effect in individuals with diabetes mellitus (Anwer etal. 2012).
Taken together, data from the studies reviewed in this
section show that Spirulina and its active compound, phycocyanin, show immunomodulatory activity. Both stimulate the production of antibodies and up- or downregulate
the expression of genes encoding IL-1, IL-2, IL-4, IL-6,
IL-10, IFN-, IL-17, and TNF-. However, different studies report regulation of different sets of cytokine genes.
This discrepancy may be due to the cell lines used, agents
used to stimulate the cells, or effects of other constituents
present in the Spirulina preparations. In addition, it may be
that the reported tumoricidal effects of Spirulina platensis
are mediated indirectly via increased production of TNF-
and IFN-. Although Spirulina appears to ameliorate
immunosenescence in older subjects, further human studies
with larger cohorts are needed to support this conclusion.
Antiinflammatory activity
The immune system is the main mediator of body inflammation, with cytokines playing a central role in initiating
inflammation. The activation of the immune system results
in the production of pro-inflammatory cytokines, such as
TNF-, IL-1, and IL-6 (Abdel-Daim etal. 2015). Up to
date, the benefits of Spirulina in building the immunity and
improving resistance to inflammation response are well
documented.
It is well known that COX-2 is the principal isoform
involved in inflammation, and induction of COX-2 is
responsible for the production of prostaglandins at the site
of inflammation (Fournier and Gordon 2000). In a human
whole blood assay (Reddy etal. 2000), phycocyanin significantly inhibited COX-2 with an IC50 value of 80nM. The
anti-inflammatory activity of phycocyanin might be due, in
part, to its selective COX-2 inhibitory effect, although its
ability to efficiently scavenge free radicals and effectively
inhibit lipid peroxidation might also be involved.
Spirulina has very promising anti-inflammatory effect
on colitis and arthritis. The anti-inflammatory effect of
phycocyanin was first reported in an experimental model of
colitis in rats (Gonzlez etal. 1999). In this study, phycocyanin (150, 200, and 300mg/kg perorally) was administered
30min before induction of colitis. Phycocyanin substantially reduced myeloperoxidase (MPO) activity which was
increased in the control colitis group after 24h. Moreover,
histopathological and ultrastructural studies showed inhibition in inflammatory cell infiltration and reduction to some

Arch Toxicol (2016) 90:18171840

extent in colonic damage in rats treated with phycocyanin.


The anti-inflammatory effect of Spirulina platensis in acetic acid-induced rat experimental colitis was also reported
by Abdel-Daim etal. (2015): Spirulina platensis significantly reduced the activities of inflammation markers such
as TNF-, IL-1, and IL-6. The anti-inflammatory activity
of Spirulina was also studied in zymosan-induced arthritis in mice (Remirez etal. 2002). Spirulina (100, 400mg/
kg, perorally) was administered to mice with arthritis for
8days. The levels of zymosan-induced -glucuronidase
were significantly reduced by Spirulina. The inflammatory reaction was substantially decreased and the cartilage
of mice treated with zymosan plus Spirulina showed well
preserved chondrocytes with normal rough endoplasmic
reticulum. Similarly, Rasool etal. (2006, 2009) evaluated
the anti-inflammatory effect of Spirulina on adjuvantinduced arthritis in mice and showed that Spirulina had
promising anti-inflammatory activity against adjuvantinduced arthritic animals. In their studies, the levels of
lysosomal enzymes, tissue marker enzymes, inflammatory
mediator TNF-, -glucuronidase, and paw volume were
significantly increased in the adjuvant-induced arthritic
mice. However, oral administration of Spirulina fusiformis
(800mg/kg/b.w.) considerably reduced the above physical and biochemical parameters to near normal conditions
(Rasool etal. 2006, 2009).
Currently, the pathogenesis of nonalcoholic steatohepatitis (NASH) remains unclear, but accumulating data suggest
that oxidative stress and the relationship between inflammation and immunity play a crucial role. In NASH model
rats (Pak etal. 2012), increases in plasma liver enzymes,
liver fibrosis, and ROS production as well as NF-B activation and changes in the lymphocyte surface antigen ratio
(CD4+/CD8+) were observed. Spirulina and phycocyanin administration significantly abated these changes and
lessen the inflammatory responses. Polysaccharide extract
from Spirulina platensis inhibited alkali burn-induced corneal neovascularization and inflammation. For example,
Yang etal. (2012) showed that Spirulina markedly inhibited inflammation by reducing both the mono- and polymorphonuclear cell infiltration into the corneal stroma and
levels of stromal cell-derived factor-1 (SDF1), TNF-, and
macrophage inflammatory protein-3 (MIP3a).
Brain inflammation plays a major role in the neurodegenerative diseases. In particular, microglial cell activation
is believed to be associated with the pathogenesis of neurodegenerative diseases, including Parkinsons disease (PD).
Spirulina reverses this effect due to its anti-inflammatory
properties. In the study carried out by Pabon etal. (2012),
an adeno-associated virus vector (AAV9) for -synuclein
was injected in the substantia nigra of PD model rats; the
effects of Spirulina on the inflammatory response were
further studied. Spirulina was neuroprotective in this

1829

-synuclein model of PD as more tyrosine hydroxylase


positive cells (TH+) and NeuN+ cells were observed. In
addition, Spirulina concomitantly decreased the number
of activated microglial cells as determined by major histocompatibility complex (MHC class II) expression. Chen
etal. (2012) also studied the anti-inflammatory effect of
Spirulina and phycocyanin in microglial cells. Their group
found that Spirulina platensis and phycocyanin significantly reduced lipopolysaccharide-induced lactate dehydrogenase release and inhibited expression of inflammationrelated genes iNOS, COX-2, TNF-, and IL-6. Another
disease, tinnitus, arises from an increase in excitatory neurotransmission and is associated with N-methyl d-aspartate
receptor (NMDA receptor, NR) activity. Pro-inflammatory
cytokines lead to tinnitus directly or via modulating NR
gene expression (Hwang etal. 2011a, b). In the study of
Hwang etal. (2013), a four-day salicylate treatment caused
a significant increase in N-methyl d-aspartate receptor subunit (2BNR2B), TNF-, and IL-1 mRNAs expression in
the cochlea and inferior colliculus of mice. Dietary supplementation with phycocyanin or Spirulina platensis water
extract significantly reduced the salicylate-induced tinnitus
and downregulated the mRNAs expression of NR2B, TNF, IL-1, and COX-2 in the cochlea and inferior colliculus
of mice.
Spirulina platensis and Spirulina lonar show antipyretic and anti-inflammatory properties. However, Spir
ulina platensis is more effective than Spirulina lonar as
anti-inflammatory and anti-pyretic agent. The anti-inflammatory and anti-pyretic properties of Spirulina platensis
and Spirulina lonar were evaluated in rat models (Somchit
etal. 2014). For anti-inflammatory activity assessment, the
reduction in the volume of paw edema induced by prostaglandin E2 was measured. Rats were dosed orally with
2 or 4mg/kg of Spirulina platensis or Spirulina lonar. A
significant dose-dependent reduction in the diameter of
paw edema was observed. Spirulina platensis treatment
revealed faster reduction in rectal temperature. Due to its
antioxidant and anti-inflammatory activities, Spirulina
slowed the development of arthritis in rats and inhibited the
development of both macroscopic and microscopic histopathological lesions in rats with Freunds adjuvant-induced
arthritis (Ali etal. 2015). In the study conducted by Yogianti etal. (2014), the inhibitory effect of dietary Spirulina
platensis on UVB-induced skin inflammatory response
was evaluated in Ogg1-knockout (KO) mice and the wildtype counterpart. Dietary Spirulina platensis suppressed
the tumor induction and development. Moreover, induction of erythema and ear swelling, one of the hallmarks
of UVB-induced inflammatory responses, was suppressed
in the skin of Ogg1-KO mice and albino hairless mice fed
with dietary Spirulina platensis. These findings suggest
that Spirulina platensis exerts anti-tumor effects against

13


1830

UVB irradiation in the skin through its anti-inflammatory


and antioxidant effects. In another study (Muga and Chao
2014), high cholesterol levels enhanced inflammation in
hamsters; however, Spirulina platensis diets regulated proinflammatory cytokines to near the control levels.
In addition to these above animal model studies, some
novel anti-inflammatory peptides were purified from the
enzymatic hydrolysate of Spirulina maxima (Vo etal.
2013). Purification of the hydrolysate resulted in two peptides with amino acid sequences of LDAVNR (P1, 686Da)
and MMLDF (P2, 655Da). Both P1 and P2 showed a suppressive effect on IL-8 expression in histamine-stimulated
EA.hy926 endothelial cells. The production of intracellular
ROS in the mast and endothelial cells was decreased by
P1 or P2. The peptides P1 and P2 from Spirulina maxima
could be used as functional ingredients with potent antiinflammatory benefits.
Taken together, from the various animal model experiments, the anti-inflammatory effects of Spirulina are an
important aspect underlying its use as a treatment for
many human and animal diseases. Under normal conditions immunomodulatory and anti-inflammatory activities
go hand in hand (Wang etal. 2014a, b; Vzquez-Velasco
etal. 2014). Due to the anti-inflammatory properties, Spirulina shows very efficient ameliorative effects on colitis
and arthritis, as well as PD. However, it is worthy to note
that all these conclusions are drawn from animal studies;
many more studies need to be performed in human subjects
if these effects are to be fully validated. Future clinical trials are required to establish the anti-inflammatory benefits
in human. The immunomodulatory and anti-inflammatory
activities of Spirulina and phycocyanin are summarized in
Table3.
Mechanism(s) ofantioxidant andimmune properties
Most research into Spirulina has focused on its antioxidant, anti-inflammatory, and immunomodulatory effects in
humans and animals. However, few studies have attempted
to identify the mechanisms underlying these effects. Thus,
our understanding in this area remains limited. Nevertheless, some studies provide preliminary information regarding the beneficial properties of Spirulina.
The first task was to analyze and identify the active
compounds in Spirulina. Spirulina contains several active
ingredients, particularly phycocyanin, phycocyanobilin,
and -carotene, all of which show promising antioxidant,
immunomodulatory, and anti-inflammatory activity (Shalaby and Shanab 2013; Mallikarjun Gouda etal. 2015).
Phycocyanin scavenges hydroxyl, alkoxyl, and peroxyl
radicals, as well as suppressing iNOS expression, reducing
nitrite production, and inhibiting lipid peroxidation in liver
microsomes (Deng and Chow 2010; Riss etal. 2007; Wang

13

Arch Toxicol (2016) 90:18171840

etal. 2007). Phycocyanin also inhibits expression of genes


that regulate critical factors involved in tumorigenesis in
mice, such as ornithine decarboxylase, COX-2, IL-6, and
pSTAT3 (Gupta and Gupta 2012). The chemical structure
of phycocyanobilin is similar to that of bilirubin (Fig.1),
which is a highly specific inhibitor of NADPH oxidase
(Lanone etal. 2005; Matsumoto etal. 2006); thus, McCarty (2007) hypothesized that phycocyanobilin would also
inhibit NADPH oxidase. Indeed, Riss etal. (2007) observed
a marked reduction in NADPH oxidase activities in hamsters that were fed phycocyanin. In addition, phycocyanin (the major antioxidant in Spirulina) might also act by
scavenging ROS (Romay etal. 2003; Gutirrez-Rebolledo
etal. 2015). Trypsin-mediated hydrolysis of phycocyanins
revealed that the apoprotein portion of the molecule makes
a significant contribution to its antioxidant activity (Zhou
etal. 2005). -Carotene, another major constituent of Spirulina, also has antioxidant and anti-inflammatory effects
(Deng and Chow 2010) and provides protection against singlet oxygen-mediated lipid peroxidation; thus, it is considered to be an efficient membrane antioxidant (Schafer etal.
2002). Indeed, the compound directly blocks the intracellular accumulation of ROS in RAW264.7 cells (Bai etal.
2005) and inhibits the expression of inflammation-associated genes (iNOS, COX-2, TNF-, and IL-1) in LPSstimulated RAW264.7 cells and LPS-treated mice. In addition, NF-B and iNOS promoter activity are also blocked
by -carotene. -Carotene blocks nuclear translocation of
the NF-B p65 subunit, which correlates with its inhibitory effects on IBa phosphorylation and degradation (Bai
etal. 2005). -Carotene accumulates in RAW264.7 cells
and induces changes in intracellular redox status, which
in turn regulate the immune function of macrophages
(Katsuura etal. 2009). Thus, these early studies show that
phycocyanin, phycocyanobilin, and -carotene are major
contributors to the antioxidant, anti-inflammatory, and
immunomodulatory activities of Spirulina.
Several studies have attempted to elucidate the signal
transduction pathways involved in the antioxidant and antiinflammatory effects of Spirulina. One study found that
phycocyanin from Spirulina platensis rapidly stimulated
phosphorylation of inflammation-related signaling molecules such as ERK, JNK, p38, and IB (Chen etal. 2014).
Moreover, our own group hypothesized that Spirulina activates the JAK/STAT signaling pathways because this pathway is downstream of the MAPK pathway (Wang etal.
2012; Wu etal. 2014a). Yogianti etal. (2014) showed that
Spirulina platensis effectively downregulates p38, JNK,
and extracellular signal-regulated kinase in mice exposed
to UVB radiation. The major active compound in Spirulina, phycocyanin, also protects pancreatic beta cells from
human islet amyloid polypeptide (hIAPP)-induced apoptosis by attenuating oxidative stress and modulating the JNK

Arch Toxicol (2016) 90:18171840

and p38 pathways (Li etal. 2009). In addition, phycocyanin regulates the p38 and ERK1/2 pathways and acts as an
antioxidant and anti-apoptotic agent to attenuate cardiac
dysfunction (Khan etal. 2006). Therefore, the up- or downregulation of MAPK, ERK, JNK, and p38 makes a significant contribution to the antioxidant and anti-inflammatory
activities of Spirulina. However, it must be noted that all of
these studies are preliminary. Further studies should examine the effects of Spirulina on key signaling pathways, such
as the JAK/STAT pathway, and their crosstalk with other
signaling events.
Although no studies have yet examined the potential
crosstalk between signaling pathways activated by Spirulina, recent studies show a close correlation between the
antioxidant and anti-inflammatory activities of this microorganism. Spirulina inhibits the inflammatory response
via both antioxidative and anti-inflammatory mechanisms,
thereby breaking the crosstalk between pathways associated with oxidative stress and those associated with inflammation, and effectively inhibiting NASH progression in a
rat model (Pak etal. 2012). Similarly, the protection against
chronic inflammation afforded by Spirulina maxima is due
to its antioxidant activity (Gutirrez-Rebolledo etal. 2015).
Spirulina platensis inhibits LPS-induced inflammatory
activity in RAW264.7 macrophages by suppressing histone
deacetylases (Pham etal. 2013). Thus, it is clear that there
is a strong relationship between the antioxidant and antiinflammatory activities induced by Spirulina.
In summary, phycocyanin and -carotene are the most
important active compounds in Spirulina and both play a
role in its antioxidant and anti-inflammatory activities. Phycocyanin scavenges free radicals, suppresses iNOS expression and nitrite production, and inhibits lipid peroxidation.
-Carotene protects against singlet oxygen-mediated lipid
peroxidation, blocks the intracellular accumulation of ROS,
and inhibits expression of inflammatory genes. Spirulina
also regulates the ERK1/2, JNK, p38, and IB signaling
pathways, resulting in antioxidant and anti-inflammatory
effects. Future research should examine other key signaling
pathways induced by Spirulina, along with their functional
crosstalk. The mechanisms thought to underlie the antioxidant, anti-inflammatory, and immunomodulatory activities
of Spirulina are presented in Fig.2.
Clinical studies
Potential clinical uses of Spirulina have been intensively
investigated. However, in contrast to the numerous animal
studies, a relative low number of clinical trials have been
carried out to evaluate the antioxidant and anti-inflammatory activities of Spirulina in humans.
To date, few studies evaluating the antioxidant properties of Spirulina have been conducted in humans.

1831

Spirulina shows favorable antioxidant effects and benefits


the immune system of healthy elderly human subjects (Park
etal. 2008). Park etal. found that Spirulina supplementation induced a significant rise in plasma IL-2 levels, while
at the same time reducing IL-6 levels. Moreover, a significant time-dependent change in antioxidant status, including an increase in SOD activity, was observed. In addition,
Spirulina led to a real clinical improvement in patients with
oral submucous fibrosis (OSF) when used as an adjuvant
antioxidant (Shetty etal. 2013).
Spirulina prevents skeletal muscle damage caused by
exercise-induced oxidative stress (Lu etal. 2006; Kalafati
etal. 2010). Sports students volunteered to take Spirulina
platensis for 3weeks, after which blood samples were
analyzed (Lu etal. 2006). The results showed that MDA
levels were significantly reduced while SOD levels were
markedly increased in those who had taken Spirulina. In
addition, the lactate concentration in the blood increased.
Taken together, these results show that ingesting Spirulina
prevents skeletal muscle damage under conditions of oxidative stress and may even extend the time taken to reach
exhaustion during exercise. A similar investigation exploring the ergogenic and antioxidant effects of Spirulina in
humans during sporting activities was performed by Kalafati etal. (2010). Moderately trained males were recruited
to a double-blind, placebo-controlled, counterbalanced
crossover study. The results showed that those who had
taken Spirulina showed a significant reduction in the rate
of carbohydrate oxidation (10.3%) during a 2-h run compared with those who had taken a placebo. In addition, the
levels of the antioxidant compound GSH were higher after
Spirulina supplementation, both at rest and at 24h after
exercise. Importantly, Spirulina attenuated the exerciseinduced increase in lipid peroxidation. However, there was
an observed increase of fat oxidation after treatment with
Spirulina. The reasons for this are still not fully understood.
COPD is closely correlated with oxidative stress.
Recently, Ismail etal. (2015) evaluated the effect of Spirulina on oxidative stress and antioxidant status in COPD
patients. The antioxidant status of COPD patients improved
after a 2-month course of Spirulina. Patients that took Spirulina for 30 and 60days, at 500mg twice per day, showed
a significant reduction in the serum levels of MDA, lipid
hydroperoxide, and cholesterol, whereas the levels of
GSH and vitamin C and SOD and GST activity increased
markedly.
In an earlier study, the effects of Spirulina platensis on
the human innate immune system were investigated (Hirahashi etal. 2002). Healthy male volunteers were orally
given 50mL/day of Spirulina platensis hot water extract
for 2months. The extract significantly increased interferon (IFN)- production from CD56-positive NK cells
in response to IL-12 and IL-12 plus IL-18 stimulation. As

13


1832

Arch Toxicol (2016) 90:18171840

Table3Immunomodulatory and anti-inflammatory activities of Spirulina and phycocyanin


Object

Specific effects

Component

References

Human peripheral blood


mononuclear cells

The secretion of IL-1, IL-4, and INF-


was enhanced

Spirulina

Mao etal. (2000)

BV-2 microglial cells

Expression of iNOS, COX-2, TNF-,


and IL-6 mRNAs was reduced

Spirulina, phycocyanin

Chen etal. (2012)

J774A.1 cells

Induced secretion of TNF-, IL-1, and


IL-6; increased pro-IL-1 and COX-2
protein expression; rapidly stimulated
phosphorylation of inflammatoryrelated signaling molecules, ERK,
JNK, p38 and I B
Decreased IL-4 production and
increased IFN- production

Phycocyanin

Chen etal. (2014)

Phycocyanin

El Sheikh etal. (2014)

Human CCD-986sk cells

TNF- and IL-6 were effectively


inhibited

Spirulina maxima

Sin etal. (2014)

Fa/Fa rat

The levels of eNOS, iNOS, and TNF-


were improved

Spirulina-surimi

Vzquez-Velasco etal. (2014)

Rat

The paw edema induced by prostaglandin E-2 was significantly decreased

Spirulina platensis

Somchit etal. (2014)

NASH model rat

The increased production of SOD,


NF-B, and the change in CD4+/
CD8+ were observed in rats of NASH,
but Spirulina and phycocyanin significantly abated these changes
TNF-a, IL-1, and IL-6 production were
inhibited

Phycocyanin and Spirulina

Pak etal. (2012)

Spirulina platensis

Abdel-Daim etal. (2015)

Serum levels of COX-2, TNF-, IL-6,


and vascular endothelial growth factor
(VEGF) were decreased
The induced MPO activity was reduced;
Inhibition in inflammatory cell infiltration and reduction to some extent in
colonic damage were observed
-Glucuronidase induced by zymosan
was significantly reduced

Spirulina platensis

Ali etal. (2015)

Phycocyanin

Gonzlez etal. (1999)

Spirulina platensis

Remirez etal. (2002)

The increase of lysosomal enzymes,


tissue marker enzymes, TNF-,
-glucuronidase, and the paw volume
was reduced significantly
Significantly inhibited the humoral
immune response, cell-mediated
immune response (delayed-type
hypersensitivity reaction), and TNF-
production
Downregulated the mRNAs expression
of NR2B, TNF-, IL-1 mRNAs, and
COX-2 genes

Spirulina

Rasool etal. (2006, 2009)

Spirulina fusiformis

Rasool and Sabina (2009)

Phycocyanin and Spirulina

Hwang etal. (2013)

The secretion of IL-4 and IL-10 in


tumor-bearing mice decreased,
whereas the levels of IL-17, TNF-,
and IFN- were increased
UVB-induced inflammatory responses
were suppressed; p38, JNK, and extracellular signal-regulated kinase were
downregulated

Spirulina platensis

Shokri etal. (2014)

Spirulina platensis

Yogianti etal. (2014)

Blood mononuclear cells

Rat with colitis


Rat with arthritis

Rats with colitis

Mice with colitis


Mice with colitis

Mice

Mice

Mice with cancer

Mice

13

Arch Toxicol (2016) 90:18171840

1833

Table3continued
Object

Specific effects

Component

References

Rats with Parkinsons


disease model

More tyrosine hydroxylase positive


cells (TH+) and NeuN+ cells were
observed; numbers of activated microglial cells as determined by major
histocompatibility complex expression
were decreased
The mRNAs expression of NR2B, TNF, IL-1, and COX-2 in the cochlea
and IC of mice was downregulated
Tumor induction and development were
suppressed; induction of erythema and
ear swelling was suppressed
A significant rise in plasma IL-2
concentration and a reduction in IL-6
concentration were observed

Spirulina

Pabon etal. (2012)

Phycocyanin or Spirulina
platensis

Hwang etal. (2013)

Spirulina platensis

Yogianti etal. (2014)

Spirulina

Park etal. (2008)

Spirulina manifested increased


indoleamine 2,3-dioxygenase (IDO)
activity and ameliorated anemia and
immunosenescence in older subjects
COX-2 was inhibited with an
IC50 value of 80nM

Spirulina

Selmi etal. (2011)

Phycocyanin

Reddy etal. (2000)

The IFN- production from CD56positive NK cells was significantly


increased
The differentiation of Th2 cells mediated was suppressed by inhibiting the
production of IL-4

Spirulina platensis

Hirahashi etal. (2002)

Spirulina

Mao etal. (2005)

Plasma MDA level was reduced significantly; plasma concentrations of total


cholesterol, LDL-cholesterol, IL-6,
and blood pressure were decreased
The values of corpuscular hemoglobin
increased; corpuscular volume and
corpuscular hemoglobin concentration
increased in male participants
A statistically significant decrease in
mean probing pocket depth and gain
in the clinical attachment level were
observed
Significant clinical improvement in
mouth opening, ulcers, erosions, and
vesicles was observed

Spirulina

Lee etal. (2008)

Spirulina

Selmi etal. (2011)

Spirulina

Mahendra etal. (2013)

Spirulina

Patil etal. (2015)

The significant increase of CD4 count


cells and significant decrease of
viral load levels of the patients were
observed

Spirulina platensis

Ngo-Matip etal. (2015)

Mice treated with salicylate

Ogg1 knockout-(KO) mice

Human (6087years old)

Human (50years old)

Human whole blood


Male human (4046years old)

Human with allergic rhinitis


(1855years old)
Human with type 2 diabetes

Human (50years or older)

Human with chronic periodontitis

Human with oral submucous


fibrosis
HIV-1 patients

both IL-12 and IL-18 receptors were not upregulated by


Spirulina platensis, the augmented IFN- production was
attributed to Spirulina platensis ability to induce the formation of the functional receptor complexes. In addition,
Spirulina platensis markedly increased IL-12 p40 unit production from PBMCs treated with BCG cell wall skeleton.
BCG cell wall skeleton is a ligand for Toll-like receptors 2
and 4 in myeloid cells. Therefore, Spirulina platensis seems

to induce Toll-like receptor-mediated IL-12 secretion from


myeloid cells (monocytes); in turn, IL-12 stimulates NK
cells to produce IFN-. Glycolipids in Spirulina platensis
could be responsible for the stimulation of Toll-like receptors. More than 4weeks of Spirulina platensis hot water
extract administration is required for an augmentation of
IFN- production by NK cells (Hirahashi etal. 2002). As
NK cells play a major role in the tumor and virus-infected

13


1834

cells surveillance (Langers etal. 2012), the ability of Spir


ulina platensis to activate NK cells might explain, at least
in part, its anti-tumor and anti-viral effects.
Allergic rhinitis is an IgE-mediated allergy. Th2-mediated immune response plays a crucial role in the initiation and progression of the disease (Radman etal. 2015).
IL-4, a Th2 cytokine, promotes Th2 cells differentiation
and stimulates B cells to produce IgE; IgE triggers mast
cell degranulation with the release of inflammatory mediators responsible for symptoms of allergic rhinitis. On the
other hand, Th1 cell cytokines such as IL-2 and IFN- limit
IL-4 effects by promoting cell-mediated immune responses
(Radman etal. 2015). The impact of a Spirulina-based
dietary supplement on patients with allergic rhinitis was
evaluated by assessing the production of cytokines IL-4,
IFN-, and IL-2 in E-mediated allergy (Mao etal. 2005).
In a randomized double-blinded crossover study versus
placebo, allergic individuals were fed daily with either placebo or Spirulina-based dietary supplement, at 1g or 2g
for 12weeks. Human peripheral blood mononuclear cells
were isolated to determine the levels of cytokines. Spirulina reduced IL-4 level produced by phytohemagglutininstimulated cells by 32%. Spirulina modulated the profile
in patients with allergic rhinitis by suppressing the differentiation of Th2 cells mediated, in part, by inhibiting the
production of IL-4. In another randomized double-blind
placebo-controlled study, administration of Spirulina plat
ensis tablets (2g/day for 6months) significantly ameliorated nasal discharge and congestion, sneezing and itching
in patients with allergic rhinitis (Cingi etal. 2008). In addition, Spirulina was reported to increase IgA level in human
saliva (Karkos etal. 2011). IgA, the major antibody in
saliva, has anti-adhesion effects but also antibacterial and
antiviral effects (Walker 2004).
Besides endothelial (von Willebrand factor) and procoagulant factors (D-dimer, PAI-1), inflammatory factors
(IL-1, IL-6, TNF-) are also involved in the development
of vascular complications in diabetes mellitus (Domingueti
etal. 2015). The effectiveness of Spirulina as functional
food in Korean patients with type 2 diabetes was evaluated (Lee etal. 2008). The subjects in Spirulina group
received 40 pills containing 0.2g of freeze-dried Spirulina
(8.0g/day) for 12weeks. The supplementation resulted in
a significant reduction in plasma MDA level. Moreover,
the subjects with higher initial total cholesterol and LDLcholesterol levels showed higher reduction in plasma concentrations of total cholesterol, LDL-cholesterol, IL-6, but
also in blood pressure. Spirulina supplementation seems
to be more effective in subjects with dyslipidemia. This
study provides the evidence for beneficial effects of Spirulina supplementation on blood lipid profiles, inflammatory
variables, and antioxidant capacity in human with type 2
diabetes.

13

Arch Toxicol (2016) 90:18171840

In elderly people, both cellular and humoral immune


responses are decreased, the former being more altered
with aging (McElhaney etal. 2016). Thus, anemia and
immunological dysfunction (i.e., immunosenescence) are
commonly found in older subjects. In the study by conducted by Selmi etal. (2011), a group of 40 volunteers
of both sexes with an age of 50years or older took Spirulina supplementation for 12weeks. After the treatment,
an increase in values of corpuscular hemoglobin in subjects of both sexes was detected. In addition, corpuscular
volume and corpuscular hemoglobin concentration also
increased in male participants. Older women appeared to
benefit more rapidly from Spirulina supplements. Similarly, the majority of subjects showed increased indoleamine 2,3-dioxygenase enzyme activity and white blood cell
count at 6 and 12weeks of Spirulina supplementation.
Thus, Spirulina may ameliorate anemia and immunosenescence in older subjects. Large human studies are needed to
evaluate the effectiveness of this supplement.
Spirulina shows promising efficacy in the treatment of
oral diseases due to its anti-inflammatory effect. Recently,
the clinical effects of Spirulina insitu gel as an adjunct
to scaling and root planning (SRP) in the treatment of
chronic periodontitis subjects was investigated (Mahendra
etal. 2013). Thirty-three sites with probing pocket depth
of 5mm were treated with SRP along with Spirulina gel
for 120days. A statistically significant decrease in mean
probing pocket depth and gain in the clinical attachment
level were observed as compared to the SRP alone group.
In this study, the efficacy of Spirulina as a local drug delivery system in the non-surgical treatment of periodontitis
without any side effects has been proved. During the treatment of chronic periodontitis, Spirulina exerted strong antiinflammatory effects closely connected with its antioxidant
activity. In another study (Patil etal. 2015), the efficacy
of Spirulina in the management of oral submucous fibrosis (OSMF) was studied. The patients receiving 500mg of
Spirulina for 3months showed significant clinical improvement in mouth opening, ulcers, erosions, and vesicles. Very
recently, Spirulina was reported to exert effective activity
against herpes labialis (Mader etal. 2016). Larger sample
sizes, with longer periods after treatment, and multi-institutional double-blind prospective studies for the assessment
of Spirulina effects are recommended to draw further conclusions on its utility in the treatment of these oral diseases.
Nevertheless, these studies shed a new light on the possible
treatment of periodontitis and OSMF with Spirulina-based
products.
The impact of daily supplementation with Spirulina on
the immune system of nave HIV-1 patients in Cameroon
was studied by Ngo-Matip etal. (2015). In this respect, a
prospective single-blind, randomized study was conducted
on 320 HIV-1 ARV-nave patients for 12months. The

Arch Toxicol (2016) 90:18171840

1835

Fig.2Proposed mechanisms underlying the antioxidant, anti-inflammatory, and immunomodulatory activities of Spirulina

patients participants received either Spirulina platensis


supplementation or local balanced diet without Spirulina
platensis. A significant increase in CD4 cells count and a
significant decrease in viral load levels of the patients who
received a supplementation of Spirulina platensis were
found after 6months of treatment. Hemoglobin level was
also significantly higher in the same group, while the fasting blood glucose concentration decreased after 12months
compared to control. However, the mechanism by which
Spirulina platensis induces these beneficial effects is not
well understood. Further studies need to be conducted on
the HIV-naive patients to elucidate this mechanism.
In summary, Spirulina shows antioxidant activity in
humans and therefore it might be a potential treatment for
COPD and skeletal muscle damage caused by exerciseinduced oxidative stress. Spirulina also shows promising
effects in the treatment of allergic rhinitis, type 2 diabetes,
anemia, immunological dysfunction, chronic periodontitis,
submucous fibrosis, and HIV-1 infection. However, these
results need to be confirmed in clinical trials based on large
cohorts.

Commercial andregulatory status


Spirulina is marketed throughout the world as food supplement or active ingredient in functional foods and beverages. Currently, Spirulina is produced in many countries,
including several African countries, Argentina, Burma,
Chile, China, Cuba, India, Israel, Japan, Mexico, Myanmar, Pakistan, Thailand, the USA and Vietnam (Small
2011). Japan, USA and the European countries are the
main importers of Spirulina powder. In China, some hospitals in Kunming have adopted Spirulina as an auxiliary
medicine, which proved to be effective in increasing the
levels of immunoglobulin A (IgA) and immunoglobulin
M (IgM) (Liu etal. 2000). By the end of 2009, the total
annual production of Spirulina in the Wulan Town, China,
surpassed 700 t (dry weight), which accounted for 80%
of the total productivity of Inner Mongolia, and 20% of
China (Lu etal. 2011).
Normally, Spirulina is marketed as powders, tablets,
and capsules which are used as nutritional supplements;
it is also added to foods and beverages (Priyadarshani and

13


1836

Rath 2012; DSLD 2009). Spirulina extracts, alone or in


combination with other herbal extracts/minerals/vitamins,
are also commercialized as nutritional supplements. For
example, Spirulina is produced in the form of lozenges
from such powder and enriched with vitamins A and C following the regulations of the importing countries. In USA,
the Food and Drug Administration (FDA) approved the
use of Spirulina extract as color additive in different foods
(chewing gum, candy, ice cream, dessert toppings/coatings, yogurts, etc.) and coating formulations in dietary supplement capsules and tablets (Electronic Code of Federal
Regulations 2016).
Under US regulations, Spirulina is legally marketed
before 1994 (DSHEA 1994). The Dietary Supplements
Information Expert Committee (DSI-EC) undertook a
safety evaluation of Spirulina (Marles etal. 2011). Following the DSI-EC review, the proposed United States Pharmacopeia and National Formulary (USPNF) monograph
for Spirulina (Spirulina platensis and Spirulina maxima)
provided specifications for the parameters of microscopic
description; limits of microcystins (1ppm) and heavy
metals (10g/g); and protein content (60%). In China,
the National Standard of Spirulina (Spirulina platensis and
Spirulina maxima) (GB/T 16919-1997) was set up in 1997
and regulated that heavy metals such as Pb, As, Cd, and
Hg should be 2.0, 0.5, 0.2, and 0.05mg/kg, respectively.
Total colonies should be 1104/g, and the mold number
should be 25/g. Spirulina (dried biomass of Arthrospira
platensis) is generally recognized as safe (GRAS) when
used as food ingredient in levels ranging from 0.5 to 3.0g/
serving (FDA 2003).
Health Canada documented that consumers can safely
use products made only from Spirulina blue-green algae
as these were found to be free of microcystins (Health
Canada 2008). As a precaution, Health Canada also recommended that the use of non-Spirulina cyanobacteria
by children be discontinued until further studies are conducted (Health Canada 2008). Since 2005, the Australian
Therapeutic Goods Administration (TGA) changed the
regulatory status of Spirulina maxima and Spirulina plat
ensis from excipient only to active ingredient (TGA
2005). In the European Union, most of the countries allow
the use of Spirulina in food supplements with or without
certain conditions (pre-marketing authorization or evaluation needed) (European Advisory Services 2007). The British Medicines and Healthcare Products Regulatory Agency
(MHRA) have not registered any adverse reports involving Spirulina (MHRA 2009). Currently, additional insight
is needed into the mechanisms of activity, short-term and
long-term effects, and potential harms profile of Spirulina
to assess the safety of the conventional usage of Spirulina
products (Marles etal. 2011).

13

Arch Toxicol (2016) 90:18171840

Conclusions
Many preclinical and clinical studies suggest that Spirulina
has antioxidant, immunomodulatory, and anti-inflammatory activities. Normally, these studies are performed in
cell lines, tissue homogenates, and animal models, meaning
that human studies are rare. Spirulina acts by activating cellular antioxidants/enzymes and by scavenging free radicals.
Notably, Spirulina exhibits enhanced antioxidant activity
when stimulated by additional factors that induce oxidative
stress. Animal studies show that Spirulina protects against
oxidative stress by inhibiting lipid peroxidation and preventing DNA damage, by scavenging free radicals, and/or
by increasing SOD and CAT activity. In addition, Spirulina
has gained attention as a promising fish food supplement
due to its antioxidant potential.
Notably, there seems to be a threshold concentration
above which Spirulina will taper off the antioxidant activity; however, more studies are needed to support this assertion. Since Spirulina is an effective antioxidant, it may be
effective against mycotoxins, such as trichothecenes and
fumonisins, which increase oxidative stress. Thus, Spirulina may be a potential treatment for mycotoxin-mediated
intoxication of animals and humans. Although studies in
humans are limited, data suggest that Spirulina prevents
damage to skeletal muscle under conditions of exerciseinduced oxidative stress. As reported in animal studies,
Spirulina ameliorates the effects of heavy metal-induced
toxicity, particularly lead-induced toxicity, which makes it
a potential treatment for heavy metal poisoning in humans.
However, more clinical studies are needed to confirm this.
In addition to its antioxidant activity, Spirulina (and
its major active compound phycocyanin) has a variety of
immunomodulatory and anti-inflammatory activities. It
stimulates the production of antibodies and up- or downregulates genes encoding cytokines (although the cytokines
affected differ depending on which study one reads). This
discrepancy may be due to the different cell lines examined, different sources of Spirulina, and the activity of
other unknown constituents present in the Spirulina preparations. Notably, the antioxidant, anti-inflammatory, and
immunomodulation activities are interrelated. Spirulina
inhibits inflammatory responses via antioxidative and antiinflammatory mechanisms, breaks the crosstalk between
pathways linked to oxidative stress and inflammation, and
effectively inhibits NASH progression.
Currently, our understanding of the functional mechanisms underlying the activities of Spirulina is limited.
We know that phycocyanin and -carotene are important
active ingredients in Spirulina and that they contribute to
its antioxidant, immunomodulatory, and anti-inflammatory
properties. Spirulina regulates the ERK1/2, JNK, p38, and

Arch Toxicol (2016) 90:18171840

IB signaling pathways to mediate its beneficial effects;


however, more research into the underlying mechanisms
is sorely needed. Since most of the conclusions reported
herein are based on animal studies and invitro models,
future clinical studies should focus on large human cohorts.
Finally, the possible interaction between Spirulina and the
intrinsic or extrinsic properties of the food stuffs to which it
is added requires more attention.
AcknowledgmentsThis work was financially supported by Yangtze Youth Talents Fund (Yangtze University, Grant No. 2015cqr19),
National Natural Science Foundation of China (81501269), the project of Excellence FIM UHK, as well as the long-term development
plan UHHK.
Compliance with ethical standards
Conflict of interest The authors declare that they have no conflict of
interest.

References
Abd El-Baky HH, El Baz FK, El-Baroty GS (2009) Enhancement of
antioxidant production in Spirulina platensis under oxidative
stress. Acta Physiol Plant 31:623631
Abdel-Daim MM, Abuzead SM, Halawa SM (2013) Protective role of
Spirulina platensis against acute deltamethrin-induced toxicity
in rats. PLoS ONE 8(9):e72991
Abdel-Daim MM, Farouk SM, Madkour FF etal (2015) Anti-inflammatory and immunomodulatory effects of Spirulina platen
sis in comparison to Dunaliella salina in acetic acid-induced
rat experimental colitis. Immunopharmacol Immunotoxicol
37(2):126139
Abdelkhalek NK, Ghazy EW, Abdel-Daim MM (2015) Pharmacodynamic interaction of Spirulina platensis and deltamethrin in
freshwater fish Nile tilapia, Oreochromis niloticus: impact on
lipid peroxidation and oxidative stress. Environ Sci Pollut Res
Int 22(4):30233031
Ali EA, Barakat BM, Hassan R (2015) Antioxidant and angiostatic
effect of Spirulina platensis suspension in complete Freunds
adjuvant-induced arthritis in rats. PLoS ONE 10(4):e0121523
Anwer R, Khursheed S, Fatma T (2012) Detection of immunoactive
insulin in Spirulina. J Appl Phycol 24:583591
Bai SK, Lee SJ, Na HJ etal (2005) beta-Carotene inhibits inflammatory gene expression in lipopolysaccharide-stimulated macrophages by suppressing redox-based NF-kappaB activation.
Exp Mol Med 37(4):323334
Banji D, Banji OJ, Pratusha NG etal (2013) Investigation on the role
of Spirulina platensis in ameliorating behavioural changes, thyroid dysfunction and oxidative stress in offspring of pregnant
rats exposed to fluoride. Food Chem 140(12):321331
Beheshtipour H, Mortazavian AM, Haratian P etal (2012) Effects of
Chlorella vulgaris and Arthrospira platensis addition on viability of probiotic bacteria in yogurt and its biochemical properties. Eur Food Res Technol 235(4):719728
Ben Abdallah Kolsi R, Ben Gara A, Chaaben R etal (2015) Anti-obesity and lipid lowering effects of Cymodocea nodosa sulphated
polysaccharide on high cholesterol-fed-rats. Arch Physiol Biochem 121(5):210217
Berg JP, Debiton E, Dumay J etal (2002) In vitro anti-inflammatory
and anti-proliferative activity of sulfolipids from the red alga
Porphyridium cruentum. J Agric Food Chem 50(21):62276232

1837
Bermejo P, Piero E, Villar M (2008) Iron-chelating ability and
antioxidant properties of phycocyanin isolated from a protean
extract of Spirulinaplatensis. Food Chem 110(2):436445
Bermejo-Bescs P, Piero-Estrada E, Villar del Fresno AM (2008)
Neuroprotection by Spirulina platensis protean extract and phycocyanin against iron-induced toxicity in SH-SY5Y neuroblastoma cells. Toxicol InVitro 22:14961502
Chamorro G, Salazar M, Pages N (1996) Dominant lethal study of
Spirulina maxima in male and female rats after short-term feeding. Phytother Res 10:2832
Chamorro G, Salazar M, Arajo KG etal (2002) Update on the pharmacology of Spirulina (Arthrospira), an unconventional food.
Arch Latinoam Nutr 52(3):232240
Chen JC, Liu KS, Yang TJ etal (2012) Spirulina and C-phycocyanin
reduce cytotoxicity and inflammation-related genes expression
of microglial cells. Nutr Neurosci 15(6):252256
Chen HW, Yang TS, Chen MJ etal (2014) Purification and immunomodulating activity of C-phycocyanin from Spirulina plat
ensis cultured using power plant flue gas. Process Biochem
49:13371344
Cherdkiatikul T, Suwanwong Y (2014) Production of the and
subunits of Spirulina Allophycocyanin and C-Phycocyanin in
Escherichia coli: a Comparative study of their antioxidant activities. J Biomol Screen 19(6):959965
Cingi C, Conk-Dalay M, Bal C etal (2008) The effects of spirulina
on allergic rhinitis. Eur Arch Otorhinolaryngol 265:12191223
Dar BA, KhaliqR JG etal (2014) Protective effects of dietary spirulina against cadmium chloride exposed histoarchitectural
changes in the liver of freshwater catfish Clarias batrachus
(Linnaeus, 1758). Indian J Fish 61(3):8387
Dartsch PC (2008) Antioxidant potential of selected Spirulina platen
sis preparations. Phytother. Res. 22(5):627633
Das UN (2007) -Linolenic acid therapy of human glioma-a review
of invitro, invivo, and clinical studies. Med Sci Monit
13(7):119131
Deng R, Chow TJ (2010) Hypolipidemic, antioxidant and antiinflammatory activities of microalgae spirulina. Cardiovasc Ther
28(4):e33e45
Ding J, Jin A, Shi L etal (2004) Effect of spirulina on antioxdation
ability of liver during CCl4 induced chronic liver injury in mice.
J Lake Sci 16(4):343348
Dohnal V, Wu Q, Kuca K (2014) Metabolism of aflatoxins: key
enzymes and interindividual as well as interspecies differences.
Arch Toxic 88:16341644
Domingueti CP, Dusse LM, Carvalho MD, etal (2015) Diabetes mellitus: the linkage between oxidative stress, inflammation, hypercoagulability and vascular complications. J Diabetes Complications. doi:10.1016/j.jdiacomp.2015.12.018. [Epub ahead of
print]
DSHEA (1994) Dietary Supplement Health and Education Act. Public Law 103-417. http://www.health.gov/dietsupp/ch1.htm.
Accessed 5 April 2016
DSLD (2009) Dietary Supplements Labels Database. http://dietarysupplements.nlm.nih.gov/dietary/index.jsp. Accessed 5 April
2016
El Baz FK, El Baroty GS, Abd El Baky HH etal (2013) Structural
characterization and biological activity of sulfolipids from
selected marine algae. Grasas Aceites 64(5):561571
El Sheikh SM, Shalaby MAM, Hafez RA etal (2014) The immunomodulatory effects of probiotic bacteria on peripheral blood
mononuclear cells (PBMCS) of allergic patients. Am J Immunol 10(3):116130
Electronic Code of Federal Regulations (2016) Title 21, Chapter I,
Subchapter A, Part 73, Subpart A, 73.530. http://www.ecfr.
gov/cgi-bin/text-idx?mc=true&tpl=true&tpl=ecfrbroowse/
Title21/21cfr73_main_02.tpl. Accessed 5 April 2016

13


1838
Elshazly MO, Abd El-Rahman SS, Morgan AM etal (2015) The
remedial efficacy of Spirulina platensis versus chromiuminduced nephrotoxicity in male sprague-dawley rats. PLoS
ONE 10(6):e0126780
El-Tantawy WH (2015) Antioxidant effects of Spirulina supplement
against lead acetate-induced hepatic injury in rats. J Traditional
Complem Med. doi:10.1016/j.jtcme.2015.02.001
European Advisory Services (2007) The use of substances with nutritional or physiological effect other than vitamins and minerals
in food supplements. Study undertaken for DG SANCO, European Comission. http://ec.europa.eu/food/safety/docs/labellingnutrition-supplements-2007_a540169_study_other_substances_
en.pdf. Accessed 5 April 2016
Farina M, Soares FA, Zeni G etal (2004) Additive prooxidative effect
of methylmercury and ebselen in liver from suckling rat pups.
Toxicol Lett 146(3):227235
FDA (2003) Agency Response Letter GRAS Notice No.GRN 000127.
http://www.fda.gov/Food/FoodIngredientsPackaging/GenerallyRecognizedasSafeGRAS/GRASListings/ucm153944.htm.
Accessed 5 April 2016
Fournier DB, Gordon GB (2000) COX-2 and colon cancer: potential
targets for chemoprevention. J Cell Biochem Suppl 34:97102
Gad AS, Khadrawy YA, El-Nekeety AA etal (2011) Antioxidant
activity and hepatoprotective effects of whey protein and Spirulina in rats. Nutrition 27(5):582589
GB/T 16919-1997. Food grade Spirulina powder. http://www.tsinfo.
js.cn/inquiry/gbtdetails.aspx?A100=GB/T%2016919-1997.
Accessed 5 April 2016
Gonzlez R, Rodrguez S, Romay C etal (1999) Anti-inflammatory
activity of phycocyanin extract in acetic acid-induced colitis in
rats. Pharmacol Res 39(1):5559
Gupta NK, Gupta KP (2012) Effects of C-Phycocyanin on the representative genes of tumor development in mouse skin exposed to
12-O-tetradecanoyl-phorbol-13-acetate. Environ Toxicol Pharmacol 34(3):941948
Gurbuz N, Coskun ZK, Omeroglu S etal (2013) Antioxidative
and therapeutic effects of spirulina on trichloroethylene
induced cutaneous irritation balb/c mice. Bratisl Lek Listy
114(4):192198
Gutirrez-Rebolledo GA, Galar-Martnez M, Garca-Rodrguez
RV, Chamorro-Cevallos GA etal (2015) Antioxidant effect
of Spirulina (Arthrospira) maxima on chronic inflammation
induced by Freunds complete adjuvant in rats. J Med Food
18(8):865871
Hassan AM, Abdel-Aziem SH, Abdel-Wahhab MA (2012) Modulation of DNA damage and alteration of gene expression during aflatoxicosis via dietary supplementation of Spirulina
(Arthrospira) and Whey protein concentrate. Ecotoxicol Environ Saf 79:294300
Health Canada (2008). Blue green algae (Cyanobacteria) and their
toxins. http://www.hc-sc.gc.ca/ewh-semt/pubs/water-eau/cyanobactereng.php. Accessed 5 April 2016
Hirahashi T, Matsumoto M, Hazeki K etal (2002) Activation of the
human innate immune system by Spirulina: augmentation of
interferon production and NK cytotoxicity by oral administration of hot water extract of Spirulina platensis. Int Immunopharmacol 2(4):423434
Hirata T, Tanaka M, Ooike M etal (1999) Radical scavenging activities of phycocyanobilin prepared from a cyanobacterium, Spir
ulina platensis. Fisheries Sci 65:971972
Hirata T, Tanaka M, Ooike M etal (2000) Antioxidant activities of
phycocyanobilin prepared from Spirulina platensis. J Appl Phycol 12:435439
Hoang MH, Kim JY, Lee JH etal (2015) Antioxidative, hypolipidemic,
and anti-inflammatory activities of sulfated polysaccharides
from Monostroma nitidum. Food Sci Biotechnol 24(1):199205

13

Arch Toxicol (2016) 90:18171840


Hoseini SM, Khosravi-Darani K, Mozafari MR (2013) Nutritional
and medical applications of spirulina microalgae. Mini Rev
Med Chem 13(8):12311237
Huang Z, Guo BJ, Wong RNS etal (2007) Characterization and antioxidant activity of selenium-containing phycocyanin isolated
from Spirulina platensis. Food Chem 100:11371143
Hwang JH, Chen JC, Yang SY etal (2011a) Expression of COX-2 and
NMDA receptor genes at the cochlea and midbrain in salicylate
induced tinnitus. Laryngoscope 121:361364
Hwang JH, Lee IT, Jeng KC etal (2011b) Spirulina prevents memory
dysfunction, reduces oxidative stress damage and augments
antioxidant activity in senescence-accelerated mice. J Nutr Sci
Vitaminol 57(2):186191
Hwang JH, Chen JC, Chan YC (2013) Effects of C-phycocyanin and
Spirulina on salicylate-induced tinnitus, expression of NMDA
receptor and inflammatory genes. PLoS ONE 8(3):e58215
Ibrahim AE, Abdel-Daim MM (2015) Modulating Effects of Spir
ulina platensis against Tilmicosin-induced cardiotoxicity in
mice. Cell J 17(1):137144
Ismail M, Hossain MF, Tanu AR etal (2015) Effect of spirulina intervention on oxidative stress, antioxidant status, and lipid profile
in chronic obstructive pulmonary disease patients. Biomed Res
Int 2015:486120
Jeyaprakash K, Chinnaswamy P (2007) Antioxidant property of Spirulina and Liv-52 against lead induced toxicity in albino rats. J
Nat Remed 7(1):8085
Kalafati M, Jamurtas AZ, Nikolaidis MG etal (2010) Ergogenic and
antioxidant effects of spirulina supplementation in humans.
Med Sci Sports Exerc 2(1):142151
Karkos PD, Leong SC, Karkos CD etal (2011) Spirulina in clinical
practice: evidence-based human applications. Evid Based Complement Alternat Med 2011:531053
Katsuura S, Imamura T, Bando N etal (2009) beta-Carotene and
beta-cryptoxanthin but not lutein evoke redox and immune
changes in RAW264 murine macrophages. Mol Nutr Food Res
53(11):13961405
Khan Z, Bhadouria P, Bisen PS (2005) Nutritional and therapeutic
potential of Spirulina. Curr Pharm Biotechnol 6(5):373379
Khan M, Varadharaj S, Ganesan LP etal (2006) C-phycocyanin
protects against ischemia-reperfusion injury of heart through
involvement of p38 MAPK and ERK signaling. Am J Physiol
Heart Circ Physiol 290(5):21362145
Kim MY, Cheong SH, Lee JH etal (2010) Spirulina improves antioxidant status by reducing oxidative stress in rabbits fed a highcholesterol diet. J Med Food 13(2):420426
Kim SS, Rahimnejad S, Kim KW etal (2013) Effects of dietary
supplementation of spirulina and quercetin on growth, innate
immune responses, disease resistance against edwardsiella
tarda, and dietary antioxidant capacity in the juvenile olive
flounder paralichthys olivaceus. Fish Aquat Sci 16(1):714
Konckov R, Vankov K, Vankov J etal (2014) Anti-cancer effects
of blue-green alga Spirulina platensis, a natural source of bilirubin-like tetrapyrrolic compounds. Ann Hepatol 13(2):273283
Kris-Etherton P, Harris W, Appel LJ (2003) Fish consumption, fish
oil, omega-3 fatty acids, and cardiovascular disease. Arterioscler Thromb Vasc Biol 23:2030
Kulshreshtha A, Zacharia AJ, Jarouliya U etal (2008) Spirulina in
health care management. Curr Pharm Biotechnol 9(5):400405
Langers I, Renoux VM, Thiry M etal (2012) Natural killer cells: role
in local tumor growth and metastasis. Biologics 6:7382
Lanone S, Bloc S, Foresti R etal (2005) Bilirubin decreases nos2 expression via inhibition of NAD(P)H oxidase: implications for protection against endotoxic shock in rats. FASEB J 19:18901892
Lee JB, Hayashi T, Hayashi K etal (1998) Further purification and
structural analysis of calcium spirulan from Spirulina platensis.
J Nat Prod 61:11011104

Arch Toxicol (2016) 90:18171840


Lee EH, Park JE, Choi YJ etal (2008) A randomized study to establish the effects of spirulina in type 2 diabetes mellitus patients.
Nutr Res Pract 2(4):295300
Li XL, Xu G, Chen T etal (2009) Phycocyanin protects INS-1E
pancreatic beta cells against human islet amyloid polypeptideinduced apoptosis through attenuating oxidative stress and
modulating JNK and p38 mitogen-activated protein kinase
pathways. Int J Biochem Cell Biol 41(7):15261535
Liu YF, Xu LZ, Cheng N etal (2000) Inhibitory effect of phycocyanin from Spirulina platensis on the growth of human leukemia
K562 cells. J Appl Phycol 12:125130
Lorenz RT (1999) A review of Spirulina and Haematococcus algae
meal as a carotenoid and vitamin supplement for poultry. Spirulina Pacifica Tech Bull 53:114
Lu HK, Hsieh CC, Hsu JJ etal (2006) Preventive effects of Spirulina
platensis on skeletal muscle damage under exercise-induced
oxidative stress. Eur J Appl Physiol 98(2):220226
Lu YM, Xiang WZ, Wen YH (2011) Spirulina (Arthrospira) industry
in Inner Mongolia of China: current status and prospects. J Appl
Phycol 23:265269
Mader J, Gallo A, Schommartz T etal (2016) Calcium spirulan
derived from Spirulina platensis inhibits herpes simplex virus 1
attachment to human keratinocytes and protects against herpes
labialis. J Allergy Clin Immunol 137(1):197203
Mahendra J, Mahendra L, Muthu J etal (2013) Clinical effects of
subgingivally delivered spirulina gel in chronic periodontitis cases: a placebo controlled clinical trial. J Clin Diagn Res
7(10):23302333
Mallikarjun Gouda KG, Udaya Sankar K, Sarada R etal (2015)
Supercritical CO2 extraction of functional compounds from
Spirulina and their biological activity. J Food Sci Technol
52(6):36273633
Mao TK, Van De Water J, Gershwin ME (2000) Effect of spirulina on
the secretion of cytokines from peripheral blood mononuclear
cells. J Med Food 3(3):135140
Mao TK, Van de Water J, Gershwin ME (2005) Effects of a Spirulinabased dietary supplement on cytokine production from allergic
rhinitis patients. J Med Food 8(1):2730
Marles RJ, Barrett ML, Barnes J etal (2011) United States pharmacopeia safety evaluation of spirulina. Crit Rev Food Sci Nutr
51(7):593604
Matsumoto H, Ishikawa K, Itabe H etal (2006) Carbon monoxide and
bilirubin from heme oxygenase-1 suppresses reactive oxygen
species generation and plasminogen activator inhibitor-1 induction. Mol Cell Biochem 291:2128
McCarty MF (2007) Clinical potential of spirulina as a source of phycocyanobilin. J Med Food 10:566570
McElhaney JE, Kuchel GA, Zhou X etal (2016) T-Cell immunity to
influenza in older adults: a pathophysiological framework for
development of more effective vaccines. Front Immunol 7:41
MHRA (2009) Medicines and Healthcare products Regulatory
Agency, UK. Data on suspected adverse drug reactions. http://
www.mhra.gov.uk//Onlineservices/Medicines/Druganalysisprints/index.htm. Accessed 5 April 2016
Miranda MS, Cintra RG, Barros SB etal (1998) Antioxidant activity of the microalga Spirulina maxima. Braz J Med Biol Res
31:10751079
Mitra S, Siddiqui WA, Khandelwal S (2015) C-Phycocyanin protects against acute tributyltin chloride neurotoxicity by modulating glial cell activity along with its anti-oxidant and antiinflammatory property: a comparative efficacy evaluation
with N-acetyl cysteine in adult rat brain. Chem Biol Interact
238:138150
Muga MA, Chao JC (2014) Effects of fish oil and spirulina on oxidative stress and inflammation in hypercholesterolemic hamsters.
BMC Complement Altern Med 14:470

1839
Nakagawa H, Montgomery WL (2007) Algae. In: Dietary supplements for the health and quality of cultured fish. Edited by
Nakagawa H, Sato S. and Gatlin III. D. CABI North American
Office Cambridge, MA 02139 USA, 133168
Ngo-Matip ME, Pieme CA, Azabji-Kenfack M etal (2015) Impact
of daily supplementation of Spirulina platensis on the immune
system of nave HIV-1 patients in Cameroon: a 12-months single blind, randomized, multicenter trial. Nutr J 14:70
Pabon MM, Jernberg JN, Morganti J etal (2012) A spirulinaenhanced diet provides neuroprotection in an -synuclein
model of Parkinsons disease. PLoS ONE 7(9):e45256
Pak W, Takayama F, Mine M, Nakamoto K etal (2012) Anti-oxidative
and anti-inflammatory effects of spirulina on rat model of nonalcoholic steatohepatitis. J Clin Biochem Nutr 51(3):227234
Park HJ, Lee YJ, Ryu HK etal (2008) A randomized double-blind,
placebo-controlled study to establish the effects of spirulina in
elderly Koreans. Ann Nutr Metab 52(4):322328
Patil S, Al-Zarea BK, Maheshwari S etal (2015) Comparative evaluation of natural antioxidants spirulina and aloe vera for the
treatment of oral submucous fibrosis. J Oral Biol Craniofac Res
5(1):1115
Pham TX, Kim B, Lee J (2013) Spirulina platensis inhibits lipopolysaccharide-induced inflammation through the repression of histone deacetylases in RAW 264.7 macrophages. FASEB J 27:1
Ponce-Canchihuamn JC, Prez-Mndez O, Hernndez-Muoz R
etal (2010) Protective effects of Spirulina maxima on hyperlipidemia and oxidative-stress induced by lead acetate in the liver
and kidney. Lipids Health Dis 9:17
Priyadarshani I, Rath B (2012) Commercial and industrial applications of micro algaea review. J Algal Biomass Utln
3(4):89100
Qing R, Ye H, Lan L, Fu H (2003) Study of the activity of two antioxidant enzymes of Spirulina maxima under excessive light stress.
J Sichuan Univ 40(3):565569
Qureshi MA, Ali RA (1996) Spirulina platensis exposure enhances
macrophage phagocytic function in cats. Immunopharmacol
Immunotoxicol 18:457463
Radman M, Golshiri A, Shamsizadeh A etal (2015) Toll-like receptor
4 plays significant roles during allergic rhinitis. Allergol Immunopathol 43(4):416420
Rasool M, Sabina EP (2009) Appraisal of immunomodulatory potential of Spirulina fusiformis: an invivo and invitro study. J Nat
Med 63(2):169175
Rasool M, Sabina EP, Lavanya B (2006) Anti-inflammatory effect of
Spirulina fusiformis on adjuvant-induced arthritis in mice. Biol
Pharm Bull 29(12):24832487
Rasool MK, Sabina EP, Nithya P, Lavanya K (2009) Suppressive
effect of Spirulina fusiformis in relation to lysosomal acid
hydrolases, lipid peroxidation, antioxidant status, and inflammatory mediator TNF-alpha on experimental gouty arthritis in
mice. Orient Pharm Exp Med 9(2):164173
Reddy CM, Bhat VB, Kiranmai G etal (2000) Selective inhibition of cyclooxygenase-2 by C-phycocyanin, a biliprotein
from Spirulina platensis. Biochem Biophys Res Commun
277(3):599603
Remirez D, Gonzlez R, Merino N etal (2002) Inhibitory effects
of Spirulina in zymosan-induced arthritis in mice. Mediators
Inflamm 11(2):7579
Riss J, Dcord K, Sutra T etal (2007) Phycobiliprotein C-phycocyanin from Spirulina platensis is powerfully responsible for
reducing oxidative stress and NADPH oxidase expression
induced by an atherogenic diet in hamsters. J Agric Food Chem
55(19):79627967
Romay C, Gonzalez R, Ledon N etal (2003) Cphycocyanin: a biliprotein with antioxidant, anti-inflammatory and neuroprotective
effects. Curr Protein Pept Sci 4:207216

13


1840
Sagara T, Nishibori N, Kishibuchi R etal (2015) Non-protein components of Arthrospira (Spirulina) platensis protect PC12
cells against iron-evoked neurotoxic injury. J Appl Phycol
27(2):849855
Sanyal AJ (2001) Nonalcoholic steatohepatitis. Indian J Gastroenterol
20:6470
Schafer FQ, Wang HP, Kelley EE etal (2002) Comparing beta-carotene, vitamin E and nitric oxide as membrane antioxidants. Biol
Chem 383(34):671681
Selmi C, Leung PS, Fischer L etal (2011) The effects of Spirulina
on anemia and immune function in senior citizens. Cell Mol
Immunol 8(3):248254
Shalaby EA, Shanab SMM (2013) Comparison of DPPH and ABTS
assays for determining antioxidant potential of water and methanol extracts of Spirulina platensis. Indian J Geo Marine Sci
42(5):556564
Sharma KM, Sharma A, Kumar A etal (2007) Spirulina fusiformis
provides protection against mercuric chloride induced oxidative
stress in Swiss albino mice. Food Chem Toxicol 45:24122419
Sharma S, Yadav N, Pandey A etal (2013) Antioxidant rich diet supplements (Spirulina and tamarind fruit pulp) mitigate hematological disorders in fluoride exposed mice. Toxicol Environ
Chem 95(10):17391747
Shetty P, Shenai P, Chatra L etal (2013) Efficacy of spirulina as an
antioxidant adjuvant to corticosteroid injection in management
of oral submucous fibrosis. Indian J Dent Res 24(3):347350
Shokri H, Khosravi A, Taghavi M (2014) Efficacy of Spirulina platen
sis on immune functions in cancer mice with systemic candidiasis. J Mycol Res 1(1):713
Sin JB, Choi WY, Lee HY (2014) Comparison of anti-inflammatory
activity of Spirulina maxima extract by ultrasonication and
water extraction process Spirulina maxima. J Korean Soc Food
Nutr 43(12):18521857
Small E (2011) Spirulina-food for the universe. Biodiversity
12(4):255265
Soheili M, Khosravi-Darani K (2011) The potential health benefits of
algae and micro algae in medicine: a review on Spirulina plat
ensis. Curr Nutr Food Sci 27(4):279285
Somchit MN, Mohamed NA, Ahmad Z etal (2014) Anti-inflammatory
and anti-pyretic properties of Spirulina platensis and Spirulina
lonar: a comparative study. Pak J Pharm Sci 27(5):12771280
Sotiroudis TG, Sotiroudis GT (2013) Health aspects of Spirulina
(Arthrospira) microalga food supplement. J Serb Chem Soc
78(3):395405
Sudharsan S, Subhapradha N, Seedevi P etal (2015) Antioxidant and
anticoagulant activity of sulfated polysaccharide from Graci
laria debilis (Forsskal). Int J Biol Macromol 81:10311038
Teng YN, Sheu MJ, Hsieh YW etal (2016) Beta-carotene reverses
multidrug resistant cancer cells by selectively modulating human P-glycoprotein function. Phytomedicine
23(3):316323
TGA (2005) Therapeutic Goods Administration, Australia. Complementary Medicin-es Evaluation Committee 52nd meeting.
August 2005. http://www.tga.gov.au/docs/pdf/cmec/cmecmi52.
pdf. Accessed 5th April 2016
Tobn-Velasco JC, Palafox-Snchez V, Mendieta L etal (2013) Antioxidant effect of Spirulina (Arthrospira) maxima in a neurotoxic model caused by 6-OHDA in the rat striatum. J Neural
Transm 120(8):11791189
Upasani CD, Balaraman R (2003) Protective effect of Spirulina on
lead induced deleterious changes in the lipid peroxidation and
endogenous antioxidants in rats. Phytother Res 17:330334

13

Arch Toxicol (2016) 90:18171840


Vzquez-Velasco M, Gonzlez-Torres L, Lpez-Gasco P etal (2014)
Liver oxidation and inflammation in Fa/Fa rats fed glucomannan/spirulina-surimi. Food Chem 159:215221
Vid J, Virsolvy A, Romain C etal (2015a) Dietary silicon-enriched
spirulina improves early atherosclerosis markers in hamsters on
a high-fat diet. Nutrition 31(9):11481154
Vid J, Romain C, Feillet-Coudray C etal (2015b) Assessment of
potential toxicological aspects of dietary exposure to siliconrich spirulina in rats. Food Chem Toxicol 80:108113
Vo TS, Ryu B, Kim SK (2013) Purification of novel anti-inflammatory peptides from enzymatic hydrolysate of the edible microalgal Spirulina maxima. J Funct foods 5:13361346
Walker DM (2004) Oral mucosal immunology: an overview. Ann
Acad Med Singapore 33(Suppl):27S30S
Wang L, Pan B, Sheng J etal (2007) Antioxidant activity of Spirulina
platensis extracts by supercritical carbon dioxide extraction.
Food Chem 105:3641
Wang X, Liu Q, Ihsan A etal (2012) JAK/STAT pathway plays a critical role in the proinflammatory gene expression and apoptosis
of RAW264.7 cells induced by trichothecenes as DON and T-2
toxin. Toxicol Sci 127:412424
Wang L, Wang X, Wu H etal (2014a) Overview on biological activities
and molecular characteristics of sulfated polysaccharides from
marine green algae in recent years. Mar Drugs 12(9):49845020
Wang Z, Wu Q, Kuca K etal (2014b) Deoxynivalenol: signaling pathways and human exposure risk assessmentan update. Arch
Toxicol 88:19151928
Wu LC, Ho JA, Shieh MC etal (2005) Antioxidant and antiproliferative activities of Spirulina and Chlorella water extracts. J Agric
Food Chem 53(10):42074212
Wu Q, Wang X, Yang W etal (2014a) Oxidative stress mediated cytotoxicity and metabolism of T-2 toxin and deoxynivalenol in animals and humans: an update. Arch Toxicol 88:13091326
Wu Q, Wang X, Wan D etal (2014b) Crosstalk of JNK1-STAT3 is
critical for RAW264.7 cell survival. Cell Signal 26:29512960
Xia D, Liu B, Luan XY etal (2016) Protective effects of C-phycocyanin on alcohol-induced acute liver injury in mice. Chin J
Oceanol Limnol 34(2):399404
Yang LL, Zhou QJ, Wang Y etal (2012) Comparison of the therapeutic effects of extracts from Spirulina platensis and amnion
membrane on inflammation-associated corneal neovascularization. Int J Ophthalmol. 5(1):3237
Yang F, Wong KH, Yang Y etal (2014) Purification and invitro antioxidant activities of tellurium-containing phycobiliproteins
from tellurium-enriched Spirulina platensis. Drug Des Devel
Ther 8:17891800
Yogianti F, Kunisada M, Nakano E etal (2014) Inhibitory effects
of dietary Spirulina platensis on UVB-induced skin inflammatory responses and carcinogenesis. J Invest Dermatol
134(10):26102619
Yoshikawa T, Naito Y (2002) What is oxidative stress. JMAJ
45(7):271276
Youn K, Lee J, Yun EY etal (2014) Biological evaluation and in silico
docking study of gamma-linolenic acid as a potential BACE1
inhibitor. J Funct Foods 10:187191
Zheng J, Inoguchi T, Sasaki S etal (2013) Phycocyanin and phycocyanobilin from Spirulina platensis protect against diabetic
nephropathy by inhibiting oxidative stress. Am J Physiol Regul
Integr Comp Physiol 304(2):R110R112
Zhou ZP, Liu LN, Chen XL etal (2005) Factors that effect antioxidant
activity of C-phycocyanins from Spirulina platensis. J Food
Biochem 29:313322