The Journal of Toxicological Sciences (J. Toxicol. Sci.

)
Vol.35, No.1, 41-47, 2010

41

Original Article

Neuroprotective effect of Centella asiatica extract (CAE) on

experimentally induced parkinsonism in aged
Sprague-Dawley rats
Nagaraja Haleagrahara1 and Kumar Ponnusamy2
Division of Human Biology, Faculty of Medicine and Health, International Medical University, No. 126, Jalan
19/155B, Bukit Jalil, 57000 Kuala Lumpur, Malaysia
2St. Matthew's University School of Medicine, Grand Cayman Islands British West Indies

(Received August 9, 2009; Accepted October 29, 2009)

ABSTRACT Reactive oxygen species (ROS) play an important role in ageing and age-related neurodegenerative changes including Parkinsons disease (PD). PD is characterized by signs of major oxidative
stress and mitochondrial damage in the pars compacta of the substantia nigra. Present study was designed
to investigate whether the Centella asiatica extract (CAE) would prevent 1-methyl-4-phenyl-1,2,3,6 - tetrahydropyridine (MPTP) - induced neurotoxicity in aged Sprague-Dawley rats. Adult, male Sprague-dawley rats of 300-350 g were divided into control, C. asiatica alone, MPTP alone (20 mg/kg, for 21 days)
and MPTP with C. asiatica (300 mg/kg for 21 days) groups. Effect of aqueous extract of C. asiatica on
oxidative biomarker levels in corpus striatum and hippocampus homogenate was examined. MPTP-chalOHQJHGUDWVHOLFLWHGDVLJQLFDQWLQFUHDVHLQOLSLGK\GURSHUR[LGHV/32SSURWHLQFDUERQ\OFRQWHQW3&&SDQG[DQWKLQHR[LGDVH;2SZKHQFRPSDUHGZLWKFRQWUROUDWV7KHUHZDV
DVLJQLFDQWGHFUHDVHLQWRWDODQWLR[LGDQWV7$SVXSHUR[LGHGLVPXWDVH62'S
JOXWDWKLRQHSHUR[LGDVH*3[SDQGFDWDODVH&$7SOHYHOVZLWK0373WUHDWPHQW6XSSOHPHQWDWLRQRI&$(UHGXFHG/32DQG3&&DQGVLJQLFDQWO\LQFUHDVHGS7$DQGDQWLR[LGDQW
HQ]\PHOHYHOVSLQFRUSXVVWULDWXPDQGKLSSRFDPSXV7KHVHUHVXOWVVKRZWKDWDGPLQLVWUDWLRQRI&
asiatica was effective in protecting the brain against neurodegenerative disorders such as Parkinsonism.
Key words: Parkinsonism, Centella asiatica, Neurotoxicity, Oxidative stress, Antioxidants
INTRODUCTION
Reactive oxygen species (ROS) has been reported to
play an important role in ageing process. Increase in ROS
production and imbalance in antioxidant defense and
repair mechanisms, leads to cell death during ageing and
age related neurological disorders (Bodis-Wollner et al.,
1991). Parkinsons disease (PD) is one of the widespread
neurodegerative diseases, caused by the loss of dopaminergic neurons in the substantia nigra of basal ganglia. PD
is characterized by resting tremor, rigidity and bradykinesia. Even though the exact mechanism of neurodegeneration in substantia nigra is elusive, alterations of several biochemical pathways appear to be involved in the
cascade of events leading to cellular dysfunction in substantia nigra. Mitochondrial dysfunction and increased
production of ROS are known to induce oxidative neuro-

degeneration in PD (Subathra et al., 2005). Though levodopa is the treatment of choice in PD, there are motor
complications and oxidative neuronal damage reported
with continuous therapy (Lan and Jiang, 1997; Savitt et
al., 2006). 1-methyl-4-phenyl-1, 2, 3, 6 - tetrahydropyridine (MPTP), when injected will be converted to the most
prominent mitochondrial complex I inhibitors, and cause
striatal dopamine depletion and loss of tyrosine hydroxylase immunoreactivity to produce Parkinsons like symptoms (Bashkatova et al., 2004; Lane et al., 2008).
Plants having medicinal properties have gained imporWDQFHQRZDGD\VEHFDXVHRIWKHLUEHQHFLDOHIIHFWRQ
human health care. Centella asiatica (Umbelliferae) syn
Hydrocotyl asiatica, belongs to the family of apiaceae
and widely found all over the world. The extract of this
plant has been used in Indian system of medicine for different ailments like asthma, wound healing, skin disor-

Correspondence: Nagaraja Haleagrahara (E-mail: hsnagaraja@gmail.com)

Vol. 35 No. 1

42
N. Haleagrahara and K. Ponnusamy

ders, ulcers and body ache (Sahu et al., 1989; Babu et

al., 1995; Suguna et al., 1996; Zainol et al., 2003; Kumar
and Gupta, 2002). The plant extract also showed central nervous system depressant activity and inhibition on
biosynthetic activity of fibroblasts (Del Vecchio et al.,
1984; Babu et al., 1995). In the recent reports, it has been
proved that C. asiatica has anti-lipid peroxidative and
free radical scavenging activities (Jayashree et al., 2003;
Gnanapragasam et al., 2004). The major active constituents present in the extract of C. asiatica extract are triterpenes and polyphenols (Inamdar et al., 1996; Zainol et
al., 2003). Triterpenes like asiatic acid and high polyphenolic contents are reported to contribute to the antioxidative activities of this plant extract (Gnanapragasam et al.,
2004).
The most important challenge in the research on PD is
to develop a neuroprotective therapy that can be supplemented with levodopa or other dopaminergic agents, in
the early course of disease to slow the progression of disease. The present study was aimed to investigate whether C.asiatica extract when administered orally would prevent MPTP induced neurotoxicity in the corpus striatum
of Sprague Dawley rats and our hypothesis was, C. asiatica extract offers neuroprotection in experimentally
induced PD.
MATERIALS AND METHODS
C. asiatica extract
Fresh whole plant of C. asiatica was procured from the
commercial market and samples of the plant were idenWLHGDQGDXWKHQWLFDWHGIRUWKHLUFRUUHFWERWDQLFDOLGHQWLty by a Botanist. The whole plants were cleaned, air-dried
and were powdered. The powder was soaked in double distilled H2O:C2H5OH (1:1) in shaking incubator at
&IRUGD\V7KHH[WUDFWHGVROXWLRQZDVOWHUHG
WKURXJK:KDWPDQ1ROWHUSDSHUDQGFRQFHQWUDWHGDQG
dried to powder.
Animals
Twenty four months old male Sprague Dawley rats
weighing 300-350 g (Conn et al., 1980; Sanchez, et al.,
2008) were obtained from Institute of Medical Research
(IMR, Kuala Lumpur, Malaysia), Kuala Lumpur and
housed under standard laboratory conditions (25 2C;
12 hr light and dark cycles). The animals were given
access to an animal diet and tap water ad libitum. The rats
were placed in polypropylene cages with three animals
per cage and were allowed to acclimatize one week prior to treatment.
Animals were randomly assigned into four groups with
Vol. 35 No. 1

eight rats per group. Control animals received saline alone.

Centella asiatica extract (CAE) alone group of rats were
administered 300 mg/kg body weight (dissolved in 0.9%
saline and administrated orally) of C. asiatica extract for
21 days (Kumar and Gupta, 2002; Gupta and Flora, 2006;
Subathra et al., 2005). MPTP alone group received 20
mg/kg body weight of MPTP (Sigma-Aldrich, St. Louis,
MO, USA) (HCl salt, dissolved in 0.9% saline, intraperitoneally), twice at 20 min interval and the rats were
maintained for 21 days (Gevaerd et al., 2001; Ramirez
et al., 2003; Kwok-Tung et al., 2008). In the fourth group,
the rats were challenged with neurotoxin-MPTP and
simultaneously administered with CAE for 21 days. All
the experimental procedures described were approved by
Institutional Research and Ethics committee.
On completion of experimental period, animals were
sacrificed by decapitation under sodium pentobarbitol anesthesia. Brain tissues were excised immediately. Regions were separated according to the method of
Glowinski and Iversen (1966) and immersed in ice cold
saline. The tissues were homogenized using 0.01 M TrisHCl buffer (pH 7.4) using glass homogenizer and superQDWDQWZDVVWRUHGDW&WLOOIXUWKHUDQDO\VLV)URPWKH
homogenate samples, lipid hydroperoxides (LPO), proteinFDUERQ\OFRQWHQW3&&[DQWKLQHR[LGDVH;2VXSHUoxide dismutase (SOD), glutathione peroxidase (GPx),
catalase (CAT) and total antioxidants (TA) were assayed
using ELISA kits (Cayman Chemicals and Pierce Biotechnology, Ann Arbor, MI, USA). The levels of enzymic and non-enzymic antioxidants, LPO, protein carbonyls
and TA were expressed as units per milligrams of proteins
of samples. Protein levels of the samples were estimated
by protein assay kits obtained from Cayman Chemicals
(Cayman Chemicals and Pierce Biotechnology).
Statistical analysis
Data were presented as Mean standard deviation. Statistical analysis was done by SPSS v 14.0 software packDJH'LIIHUHQFHVDPRQJYDULRXVJURXSVDQGWKHVLJQLcance were calculated by non-parametric Kruskal Wallis
H and Mann- Whitneys U-Tests. P value less than 0.05
ZDVFRQVLGHUHGVWDWLVWLFDOO\VLJQLFDQW
RESULTS
LPO
Corpus striatum and hippocampus lipid hydroperoxide
OHYHOVLQFUHDVHGVLJQLFDQWO\LQ0373WUHDWPHQWJURXSV
FRPSDUHG WR FRQWURO JURXSV 3  $ VLJQLILFDQW
decrease in serum LPO was seen with C. asiatica alone
WUHDWHGJURXSV3,QJURXSVWUHDWHGZLWK&$(

43
Effect of Centella asiatica extract on Parkinsonism in rats

DORQJZLWK0373WKHUHZDVDVLJQLFDQWGHFUHDVH3
LQ/32OHYHOVDQGWKLVZDVVLJQLFDQWO\OHVVWKDQ
WKHFRQWUROJURXSV37DEOHVDQG

GHFUHDVHGWRFRQWUROOHYHOV37UHDWPHQWZLWK

CAE alone showed significant increase in antioxidant
HQ]\PHOHYHOV37DEOHVDQG

PCC
A significant increase in striatum and hippocampus
3&&ZHUHUHFRUGHGDIWHU0373WUHDWPHQW3
The level of protein carbonyl was decreased in CAE with
0373JURXSV3(YHQWKRXJKWKHOHYHORI3&&
ZHUHVLJQLFDQWO\UHGXFHGZLWK&$(WUHDWPHQWLQ0373
treated rats, it did not significantly decrease below the
control levels both in striatum and hippocampus samples
(Tables 1 and 2).

SOD
6HUXP62'GHFUHDVHGVLJQLFDQWO\LQVWULDWXPDIWHU
H[SHULPHQWDOO\LQGXFHG3DUNLQVRQLVP3EXW
WUHDWPHQWZLWK&$(VLJQLFDQWO\LQFUHDVHGWKH62'OHYHOVDIWHUGD\V37DEOH6LPLODUFKDQJHV
were observed in SOD levels in hippocampus homogenates. More significant decrease in hippocampus SOD
OHYHOZDVREVHUYHGDIWHU03733DQGFRQFXUUHQW
treatment with C. asiaticaVLJQLFDQWO\HOHYDWHGWKH62'
levels (Table 2).

XO
7KHUHZDVDVLJQLILFDQWLQFUHDVHLQ;2OHYHOVERWK
in striatum and hippocampus after 21 days exposure
WR037337UHDWPHQWZLWKC. asiatica VLJQLcantly reduced these changes and the enzyme level was

GPx
CAE alone when injected at a dose of 300 mg/kg sigQLILFDQWO\LQFUHDVHGWKHFRUSXVVWULDWDO*3[OHYHOV3
0.05). But MPTP treatment decreased the GPx levels sig-

Table 1. Effect of CAE on antioxidant status in the corpus striatum of rats

Control

C. asiatica

MPTP

LPO (nmol/mg of protein)

3.17 0.8

2.76 0.09*

4.56 0.03**

2.98 0.05 *

PCC (nmol/mg protein)

1.68 0.09

1.54 0.08

2.48 0.09

1.65 0.05

;20PJSURWHLQ

3.23 0.03

2.61 0.07

4.89 0.04**

3.81 0.03

SOD (units/mg of protein)

18.20 0.92

22.54 0.86*

14.52 0.97***

17.17 1.74

&$70PJRISURWHLQV

45.28 1.90

53.71 1.12

32.20 1.74

41.73 1.23

GPx (nmol/min/mg of protein)

21.65 1.17

26.50 0.96*

16.30 1.14**

24.12 1.34

3.22 0.08

3.45 0.5

1.52 0.09***

2.97 0.09

Parameters

TA (mM/mg of protein)

C. asiatica + MPTP
**

***

Results are expressed as means S.D. of eight rats per group.

*P < 0.05; **P < 0.01; ***P < 0.001 - Control with other groups.
P < 0.05; P < 0.01; P < 0.001 - MPTP with C. asiatica + MPTP.

Table 2. Effect of CAE on antioxidant status in the hippocampus of rats

Control

C. asiatica

MPTP

LPO (nmol/mg of protein)

2.96 0.03

2.42 0.24

3.56 0.08**

2.67 0.27

PCC (nmol/mg protein)

1.76 0.07

1.69 0.18

2.26 0.18

**

1.52 0.15

;20PJSURWHLQ

3.82 0.09

2.79 0.18

5.12 0.19**

3.17 0.21

SOD (units/mg of protein)

14.50 0.32

17.57 0.13*

11.75 0.87***

16.62 1.88

&$70PJRISURWHLQV

47.76 1.74

56.24 1.30

38.52 1.82

46.77 1.92

GPx (nmol/min/mg of protein)

19.35 1.57

24.66 0.98*

14.40 1.08**

21.82 1.73

3.47 0.12

3.09 0.25

1.74 0.05***

2.65 0.16

Parameters

TA (mM/mg of protein)

**

C. asiatica + MPTP

***

Results are expressed as means S.D. of eight rats per group.

*P < 0.05; **P < 0.01; ***P < 0.001 - Control with other groups.
P < 0.05; P < 0.01; P < 0.001 - MPTP with C. asiatica + MPTP.
Vol. 35 No. 1

44
N. Haleagrahara and K. Ponnusamy

QLFDQWO\3DQGWUHDWPHQWZLWK&$(DORQJZLWK

MPTP was able to increase the serum GPx levels. Serum
GPx level reached near normal in Parkinsons group
with C. asiatica37DEOH$VLPLODUWUHQGZDV
REVHUYHGLQWKH*3[OHYHOLQKLSSRFDPSXVRIUDWV3
0.01) (Table 2).
CAT
7KHUHZDVVWDWLVWLFDOO\VLJQLFDQWGHFUHDVHLQVWULDWDO
&$7OHYHOVDIWHU0373WUHDWPHQWIRUGD\V3
&$(DORQHGLGDOVREULQJDERXWDVLJQLFDQWFKDQJH3
0.01) in serum CAT levels. But CAE administered for 21
days along with MPTP was able to increase the CAT levHOVPRUHWKDQ0373DORQH3DQGWKH&$7OHYel reached more than control group (Table 1). Hippocampus CAT level also significantly decreased after MPTP
IRUGD\V3C. asiatica reversed this change
FDXVHGE\0373DV&$7OHYHOZDVIRXQGWREHVLJQLFDQWO\KLJKHULQKLSSRFDPSXV37DEOH
TA
7KHUHZDVDVLJQLFDQWGHFUHDVHLQKLSSRFDPSXVDQG
VWULDWXP7$DIWHUH[SRVXUHRIUDWVWR03733
&RQFXUUHQWWUHDWPHQWZLWK&$(VLJQLFDQWO\LQFUHDVHG
VHUXPWRWDODQWLR[LGDQWOHYHOV30373WUHDWPHQW
IRUGD\VVLJQLFDQWO\GHFUHDVHGWKHKLSSRFDPSXV7$
3DQG&$(WUHDWPHQWZDVDEOHWRLQFUHDVHWKH
DQWLR[LGDQWFRQWHQWVDIWHUGD\V37DEOHV
and 2).
DISCUSSION
In the present study, adult Sprague Dawley rats chalOHQJHGZLWK0373VKRZHGKLJKO\VLJQLFDQWLQFUHDVHLQ
/32SURWHLQFDUERQ\OFRQWHQWVDQG;2OHYHOVLQFRUSXV
VWULDWXPDQGKLSSRFDPSXV7KHUHZDVDOVRDVLJQLFDQW
decrease in TA levels with MPTP treatment. The differences in the levels of LPO products observed in various
brain regions may be attributed to the differences in their
iron content and diverse metabolism, which influenced
the generation of ROS. Certain brain regions like striatum and hippocampus are highly enriched with non-heme
iron, which is catalytically involved in the production of
ROS (Hill and Switzer, 1984). Exposure to MPTP might
have lead to the peroxidation of membrane lipids evenWXDOO\OHDGLQJWRORVVRIPHPEUDQHLQWHJULW\DQGQDOO\
to cell death in these brain regions. MPTP is believed to
induce selective toxicity at central dopaminergic neurons
via the end products of its oxidation, 1-methyl-4-phenylpyridinium (MPP+) which inhibits oxidative metabolism at complex I of the mitochondrial respiratory chain
Vol. 35 No. 1

E\LWVVSHFLFLQKLELWLRQRI1$'+XELTXLQRQHR[LGRUHductase (Ebadi et al., 2001). Although the mechanism by

which it induces selective dopaminergic cell death has
not been fully elucidated, it was described that MPTP acts
as an inhibitor of complex I of mitochondrial respiratory pathway and produces a decrease in tissue ATP content
and increases ROS formation (Lane et al., 2008). Malonised dopaminergic neurones have been reported to be
more susceptible to neurodegeneration in PD and MPTP
toxicity (Iczkiewicz et al., 2006).
During oxidative stress in the neuronal cells there
is an increase in intracellular Ca2+ levels in the brain
(Annunziato et al., 2003). This increased intracellular Ca2+
can induce the irreversible conversion of xanthine dehyGURJHQDVH;'+WR;2ZKLFKLQWXUQFDWDO\]HVWKHR[Ldation of xanthine to provide a source of O-2. In addition,
auto-oxidation of dopamine in brain could also serve as a
source of superoxide anion (Olanow, 1993). These mechanisms could be the main reasons for the increased levHORI;2DQGUHGXFWLRQLQDFWLYLW\RI62'OHDGLQJWRDQ
overload of oxygen radicals and repression of antioxidant
enzymes with MPTP exposure.
Dysfunction of cholinergic neuronal activities has been
considered to be a cause of age-related impairment of
brain function among animal species, including human
beings. It has been documented that mitochondrial complex I activity is decreased in the substantia nigra of PD
patients (Lestienne et al., 1990; Gadaleta et al., 1990).
Studies on rodent brains have indicated age-related reducWLRQVLQFKROLQHUJLFDFWLYLWLHVVXFKDVKLJKDIQLW\FKROLQH
uptake, acetylcholine synthesis and acetyl choline release
(Curti et al., 1989; Vogelsberg et al., 1997; Rodrigues et
al., 2007). The reduced acetylcholine release is thought
to be directly related to a decreased synaptic transmission in cholinergic neurons. The acetylcholine release
per synapse was later shown to be decreased in aged rats
in experiments using synaptosomes (Rylett et al., 1993).
Oxidation of L-3,4-dihydroxy phenylalanine (L-DOPA)
and dopamine (DA) to generate semiquinones / quinones,
oxygen radicals, and other ROS may play a vital role in
neuronal cell death in PD. In particular, semiquinones /
quinones can form conjugates with thiol compounds such
as GSH and cysteine. Exposure of L-DOPA, DA, and other catacholamines to a system generating O2- radical led
to O2- -dependent depletion of added GSH (or cysteine),
accompanied by the formation of thiol-DA or thiol-DOPA
adducts. The substantia nigra of rodents, primates and
humans contain lower levels of glutathione than other brain regions (Migdalis et al., 2001). The concentrations of GPx, CAT and SOD and the TA were found to
EHGHFUHDVHGVLJQLFDQWO\LQWKHVWULDWXPDQGKLSSRFDP-

45
Effect of Centella asiatica extract on Parkinsonism in rats

pus of MPTP treated rats as compared to the control rats.

The present study showed that CAE administration
UHGXFHG/32VLJQLFDQWO\LQDJHGUDWVDQGDJHGUDWVFKDOlenged with MPTP. The reduction in LPO levels may be
due to the electron and H+ donating capacity of polyphenols present in CAE, which is attributed the termination
of LPO chain reaction based on their reducing power.
6HYHUDOVWXGLHVKDYHVKRZQWKDWDYRQRLGVLQWHUDFWZLWK
cell membranes, improving their stability, thereby protecting them from LPO (Saija et al., 1995). Antioxidant
property of CAE may possibly be attributed to the nitrigenomic phenolic compounds present which are also effective hydrogen donors, which makes them good antioxidants (Rice-Evans et al., 1995). A dose of 300 mg/kg of
C. asiatica extract has decreased the LPO and protein carbonyls, which are the products of lipid peroxidation and
a measure of free radical production. There was a significant increase in SOD levels, which is the only enzyme
using the super oxide anions as a substrate and producing the hydrogen peroxide as metabolite, with C. asiatiFDWUHDWPHQW7KHUHZDVDOVRDVLJQLFDQWLQFUHDVHLQ*3[
levels, which protects the cells and neurons against free
radicals induced damage. C. asiatica treatment produced
a significant increase in the CAT levels, compared to
MPTP alone treated group, demonstrating that the extract
of C. asiatica scavenges the hydrogen peroxide generated
by SOD (Schulz et al., 2000; Kumar and Gupta, 2002).
Thus, our study has shown that the C asiatica extract has
mitigated MPTP induced peroxide production in striatum
and hippocampus and reversed the oxidative damage, as
evidenced by increased TA and antioxidant enzyme levels in these regions.
The active constituents present in the C. asiatica extract
are triterpens like asiatic acid and asiaticoside. Different parts of the C. asiatica were also found to have high
level of phenolic contents. Mandel et al. (2006) reported
that phenolic compounds acts as a potent iron-chelating,
nitrigenomic antioxidants. The phenolic compounds such
as quercetin and catechins present in the CAE may have
different functional property such as scavenging of ROS
(Hatano et al., 1989; Zainol et al., 2003), inhibition of
the generation of free radicals and chain-breaking activity (Laranjinha et al., 1995). They may act as hydrogendonating radical scavengers by scavenging lipid alkoxyl
and peroxyl radical and spares overall antioxidant status
(Bors et al6WXGLHVKDYHGRFXPHQWHGWKDWELRavanoids and triterpenes exerts free radical scavenging,
membrane stabilizing, iron-chelating, lipotrophic, vasoacWLYHLPPXQRPRGXODWRU\DQWLPXWDJHQLF;2LQKLELWRry, anti-apoptotic, anti-ageing and neuroprotective effects
and increases cytoprotective heat shock proteins (Lahouel

et al., 2006; Broncel et al., 2007; Greenrod et al., 2005;

Kalfon et al., 2007; Mo et al., 2007; Luangaram et al.,
2007). Thus the observed free radical scavenging and
antioxidant effect of C asiatica in the present study could
be attributed to the triterpenes and phenolic content of the
whole plant.
In conclusion, the present study implies that Centella
asiatica enriched with polyphenols and triterpenes may
be particularly useful xenobiotics detoxifying agents as it
could decrease lipid peroxidation and enhance brain antiR[LGDQWVDQGVLJQLFDQWO\SUHYHQWWKHEUDLQIURPQHXURtoxic effects of MPTP. Therefore, C. asiatica could offer
a useful support to the Parkinsonism therapy by acting as
a neuroprotective antioxidant and thus prevent the neuronal damage in brain regions associated with ParkinsonLVP7KXV&$(H[WUDFWHQULFKHGZLWKELRDYDQRLGVDQG
triterpenes may be considered as a powerful neuroprotective agent for membrane molecular medicine. Further
studies may be necessary to elucidate the exact molecular
mechanisms of actions of the various constituents in CAE
against MPTP induced neurotoxicity.
ACKNOWLEDGMENT
This work was supported by a grant-in aid for Scientific Research, from International Medical University (IMU
Research Grant No: 115/2006), Kuala Lumpur, Malaysia.
REFERENCES
Annunziato, L., Amoroso, S., Pannaccione, A., Cataldi, M., Pignataro,
G., DAlessio, A., Sirabella, R., Secondo, A., Sibaud, L. and Di
Renzo, G.F. (2003): Apoptosis induced in neuronal cells by oxidative stress: role played by caspases and intracellular calcium
ions. Toxicol. Lett., 139, 125-133.
Babu, T.D., Kuttan, G. and Padikkala, J. (1995): Cytotoxic and antitumour properties of certain taxa of Umbelliferae with special
reference to Centella asiatica (L.) Urban. J. Ethnopharmacol.,
48, 53-57.
Bashkatova, V., Alam, M., Vanin, A. and Schmidt, W.J. (2004):
Chronic administration of rotenone increases levels of nitric
oxide and lipid peroxidation products in rat brain. Exp. Neurol.,
186, 235-241.
Bodis-Wollner, I., Chung, E., Ghilardi, M.F., Glover, A., Onofrj,
M., Pasik, P. and Samson, Y. (1991): Acetyl-Levo-carnitine
protects against MPTP-induced Parkinsonism in primates. J.
Neural. Transm. Park. Dis. Dement. Sect., 3, 63-72.
Bors, W., Heller, W., Michel, C. and Saran, M. (1990): Flavonoids
DVDQWLR[LGDQWVGHWHUPLQDWLRQRIUDGLFDOVFDYHQJLQJHIFLHQFLHV
Methods Enzymol., 186, 343-355.
Broncel, M., Franiak, I., Koter-Michalak, M., Duchnowicz, P. and
Chojnowska-Jezierska, J. (2007): The comparison in vitro the
effects of pravastatin and quercetin on the selected structural
parameters of membrane erythrocytes from patients with hypercholesterolemia. Pol. Merkur Lekarski., 22, 112-116.

Vol. 35 No. 1

46
N. Haleagrahara and K. Ponnusamy
Conn, P.M., Cooper, R., McNamara, C., Rogers, D.C. and
Shoenhardt, L. (1980): Qualitative change in gonadotropin during normal aging in the male rat. Endocrinology., 106, 15491553.
Curti, D., Dagani, F., Galmozzi, M.R. and Marzatico, F. (1989):
Effect of aging and acetyl-L-carnitine on energetic and cholinergic metabolism in rat brain regions. Mech. Ageing Dev., 47,
39-45.
Del Vecchio, A., Senni, I., Cossu, G. and Molinaro, M. (1984):
(IIHFWRI&HQWHOODDVLDWLFDRQWKHELRV\QWKHWLFDFWLYLW\RIEURElasts in culture. Farmacie Edition., 39, 355-364.
Ebadi, M., Govitrapong, P., Sharma, S., Muralikrishnan, D.,
Shavali, S., Pellett, L., Schafer, R., Albano, C. and Eken, J.
(2001): Ubiquinone (coenzyme q10) and mitochondria in oxidative stress of parkinsons disease. Biol. Signals Recept., 10, 224253.
Gadaleta, M.N., Petruzzella, M., Renis, F., Fracasso, F. and
Antatore, P. (1990): Reduced mitochondrial DNA transcription
in two brain regions of senescent rats: Effect of acetyl-L-carnitine. In: Structure, function and biogenesis of energy transfer
system. (Quazliariello, S., Papa, F., Palmieri, S. C., ed.), pp.135138, Elsevier, Amsterdam.
Gevaerd, M.S., Miyoshi, E., Silveira, R., Canteras, N.S., Takahashi,
R,N. and Da Cunha, C. (2001): L-Dopa restores striatal
GRSDPLQHOHYHOEXWIDLOVWRUHYHUVH0373LQGXFHGPHPRU\GHcits in rats. Int. J. Neuropsychopharmacol., 4, 361-370.
Glowinski, J. and Iversen, L.L. (1966): Regional studies of catecholamines in the rat brain. I. The disposition of [3H] norepinephrine, [3H] dopamine and [3H] dopa in various regions of the
brain. J. Neurochem., 13, 655-669.
Gnanapragasam, A., Ebenezar, K.K., Sathish, V., Govindaraju, P.
and Devaki, T. (2004): Protective effect of Centella asiatica on
antioxidant tissue defense system against adriamycin induced
cardiomyopathy in rats. Life Sci., 76, 585-597.
Greenrod, W., Stockley, C.S., Burcham, P., Abbey, M. and Fenech,
M. (2005): Moderate acute intake of de-alcoholized red wine,
but not alcohol, is protective against radiation-induced DNA
damage ex vivo-results of a comparative in vivo intervention
study in younger men. Mutat. Res., 591, 290-301.
Gupta, R. and Flora, S.J. (2006): Effect of Centella asiatica on
arsenic induced oxidative stress and metal distribution in rats. J.
Appl. Toxicol., 26, 213-222.
Hatano, T., Edamatsu, R., Hiramatsu, M., Moti, A., Fujita, Y.,
Yasuhara, T., Yoshida, T. and Okuda, T. (1989): Effects of tannins and related polyphenols on superoxide anion radical, and on
1,1-diphenyl-2-picrylhydrazyl radical. Chem. Pharmaceut. Bull.,
37, 2016-2021.
Hill, J.M. and Switzer, R.C. (1984): The regional distribution and
cellular localization of iron in the rat brain. Neuroscience., 11,
595-603.
Iczkiewicz, J., Jackson, M.J., Smith, L.A., Rose, S. and Jenner, P.
(2006): Osteopontin expression in substantia nigra in MPTPtreated primates and in Parkinsons disease. Brain Res., 1118,
239-250.
Inamdar, P.K., Yeole, R.D., Ghogare, A.B. and de Souza, N.J.
(1996): Determination of biologically active constituents in Centella asiatica. J. Chromatogr. A., 742, 127-130.
Jayashree, G., Kurup, M.G., Sudarslal, S. and Jacob, V.B. (2003):
Anti-oxidant activity of Centella asiatica on lymphoma-bearing
mice. Fitoterapia, 74, 431-434.
Kalfon, L., Youdim, M.B. and Mandel, S.A. (2007): Green tea
polyphenol (-)-epigallocatechin-3-gallate promotes the rapid
Vol. 35 No. 1

protein kinase C- and proteasome-mediated degradation of Bad:

implications for neuroprotection. J. Neurochem., 100, 992-1002.
Kumar, M.H.V. and Gupta, Y.K. (2002): Effect of different extracts
of Centella asiatica on cognition and markers of oxidative stress
in rats. J. Ethnopharmacol., 79, 253-260.
Lu, K.T., Ko, M.C., Chen, B.Y., Huang, J.C., Hsieh, C.W., Lee,
M.C., Chiou, R.Y., Wung, B.S., Peng, C.H. and Yang, Y,L.
(2008): Neuroprotective effects of resveratrol on MPTP-induced
neuron loss mediated by free radical scavenging. J. Agric. Food
Chem., 56, 6910-6913.
Lahouel, M., Amedah, S., Zellagui, A., Touil, A., Rhouati, S.,
Benyache. F., Leghouchi, E. and Bousseboua, H. (2006): The
LQWHUDFWLRQRIQHZSODQWDYRQRLGVZLWKUDWOLYHUPLWRFKRQGULD
UHODWLRQEHWZHHQWKHDQWLDQGSURR[\GDQWHIIHFWDQGDYRQRLGV
concentration. Therapie., 61, 347-355.
Lan, J. and Jiang, D.H. (1997): Excessive iron accumulation in the
brain: a possible potential risk of neurodegeneration in Parkinsons disease. J. Neural. Transm., 104, 649-660.
Lane, E.L., Carlsson, T., Kirik, D. and Dunnett, S.B. (2008): Animal models of Parkinsonism. In: Sourcebook of models for biomedical research. (Conn, P. M., ed.), pp313-320, Humana Press
Inc., Totowa, NJ.
Laranjinha, J., Vieira, O., Madeira, V. and Almedia, L. (1995): Two
related phenolic antioxidants with opposite effects on vitamin E
content in low density lipoproteins oxidized by ferrylmyoglobin: consumption vs regeneration. Arch. Biochem. Biophys., 323,
373-381.
Lestienne, P., Nelson, J., Riederer P., Jellinger K. and Reichmann,
H. (1990): Normal mitochondrial genome in brain from patients
with Parkinsons disease and complex I defect. J. Neurochem.,
55, 1810-1812.
Luangaram, S., Kukongviriyapan, U., Pakdeechote, P.,
Kukongviriyapan, V. and Pannangpetch, P. (2007): Protective
effects of quercetin against phenylhydrazine-induced vascular
dysfunction and oxidative stress in rats. Food Chem. Toxicol.,
45, 448-455.
Mandel, S., Amit, T., Reznichenko, L., Weinreb, O. and Youdim,
M.B. (2006): Green tea catechins as brain-permeable, natural
iron chelators-antioxidants for the treatment of neurodegenerative disorders. Mol. Nutr. Food Res., 50, 229-234.
0LJGDOLV,1.DORJHURSRXORX.,OLRSRXORX97ULDQWDORX3
Charalabides, J., Mortzos, G. and Cordopatis, C. (2001): Plasma levels of endothelin, lipid peroxides and prostacyclin in diabetic patients with macroangiopathy. Diabetes Res. Clin. Pract.,
54, 129-136.
Mo, S.F., Zhou, F., Lv, Y.Z., Hu, Q.H., Zhang, D.M. and Kong, L.D.
(2007): Hypouricemic action of selected flavonoids in mice:
structure-activity relationships. Biol. Pharm. Bull., 30, 15511556.
Olanow, C.W. (1993): A radical hypothesis for neurodegeneration.
Trends Neurosci., 16, 439-444.
5DPLUH]$'/LX;DQG0HQQLWL)65HSHDWHGHVWUDGLRO
treatment prevents MPTP-induced dopamine depletion in male
mice. Neuroendocrinol., 77, 223-231.
Rice-Evans, C.A., Miller, N.J., Bolwell, P.G., Bramley, P.M. and
Pridham, J.B. (1995): The relative antioxidant activities of plant
GHULYHGSRO\SKHQROLFDYRQRLGV)UHH5DGLF5HV22, 375-383.
Rodrigues, R.J., Canas, P.M., Lopes, L.V., Oliveira, C.R. and Cunha,
5$0RGLFDWLRQRIDGHQRVLQHPRGXODWLRQRIDFHW\OFKRline release in the hippocampus of aged rats. Neurobiol. Aging,
29, 1597-1601.
Rylett, R.J., Goddard, S., Schmidt, B.M. and Williams, L.R. (1993):

47
Effect of Centella asiatica extract on Parkinsonism in rats
Acetylcholine synthesis and release following continuous intracerebral administration of NGF in adult and aged Fischer-344
rats. J. Neurosci., 13, 3956-3963.
Sahu, N.P., Roy, S.K. and Mahato, S.B. (1989): Spectroscopic determination of structures of triterpenoid trisaccharides from Centella asiatica. Phytochem., 28, 2852-2854.
Saija, A., Scalese, M., Lanza, M., Marzullo, D., Bonina, F. and
Castelli, F. (1995): Flavonoids as antioxidant agents: importance
of their interactions with biomembranes. Free Radic. Biol. Med.,
19, 481-486.
Sanchez, H.L., Silva, L.B., Portiansky, E.L., Herenu, C.B., Goya,
R.G. and Zuccolilli, G.O. (2008): Dopaminergic mesencephalic systems and behavioral performance in very old rats.
Neuroscience., 154, 1598-1606.
Savitt, J.M., Dawson, V.L. and Dawson, T.M. (2006): Diagnosis and
treatment of Parkinson disease:molecules to medicine. J. Clin.
Invest., 116, 1744-1754.

Schulz, J.B., Lindenau, J., Seyfried, J. and Dichgans, J. (2000):

Glutathione, oxidative stress and neurodegeneration. Eur. J.
Biochem., 267, 4904-4911.
Subathra, M., Shila, S., Devi, M.A. and Panneerselvam, C. (2005):
Emerging role of Centella asiatica in improving age-related neurological antioxidant status. Exp. Gerontol., 40, 707-715.
Suguna, L., Sivakumar, P. and Chandrakasan, G. (1996): Effect of
Centella asiatica extract on dermal wound healing in rats. Indian
J. Exp. Biol., 34, 1208-1211.
Vogelsberg, V., Fong, T.G., Neff, N.H. and Hadjiconstantinou, M.
&KROLQHUJLFGHFLWVLQDJHGUDWVSLQDOFRUGUHVWRUDWLRQ
by GM1 ganglioside. Brain Res., 761, 250-256.
Zainol, M.K., Abd-Hamid, A., Yusof, S. and Muse, R. (2003): Antioxidant activity and total phenolic compounds of leaf, root and
petiole of four accessions of Centella asiatica (L.) Urban. Food
Chem., 81, 575-581.

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