Received: 1 January 2018 Revised: 16 May 2018 Accepted: 21 May 2018

DOI: 10.1002/ptr.6127

RESEARCH ARTICLE

Protective effects of cinnamon bark extract against ischemia–

reperfusion injury and arrhythmias in rat
Mehrnoosh Sedighi1,2 | Afshin Nazari3 | Mahdieh Faghihi4 | Mahmoud Rafieian‐Kopaei5 |

Arash Karimi6 |
Maryam Seyyed AliMoghimian7 | Mozaffarpur8 | Marzieh Rashidipour1 |
9 | 9 |
Mehrdad Namdari Mostafa Cheraghi Bahram Rasoulian3

1
Razi Herbal Medicines Research Center,
Lorestan University of Medical Sciences, Cinnamomum zeylanicum (cinnamon) is a plant with potent antioxidant activity and has
Khorramabad, Iran
been used in traditional medicine for improvement of heart function. The effects of cinna-
2
Student Research Committee, Lorestan
University of Medical Sciences, Khorramabad, Iran mon bark ethanolic extract were investigated against ischemia‐induced arrhythmias and
3
Razi Herbal Medicines Research Center and heart injury in an in vivo rat model of regional heart ischemia. The extract was also stan-
Department of Physiology, Lorestan University
dardized, and its antioxidant activity was evaluated. Adult male Sprague–Dawley rats were
of Medical Science, Khorramabad, Iran
4
Department of Physiology, School of
subjected to 30 min of ischemia by occlusion of the left anterior descending coronary
Medicine, Tehran University of Medical artery followed by 5 days of reperfusion. Thirty‐two animals were randomized to receive
Science, Tehran, Iran
5
daily oral administration of vehicle or C. zeylanicum bark extract (intragastric, 50, 100, or
Medical Plants Research Center, Basic Health
Sciences Institute, Shahrekord University of 200 mg/kg) 14 days before ischemia. C. zeylanicum was standardized through HPLC anal-
Medical Sciences, Shahrekord, Iran ysis. Administration of cinnamon bark extract significantly improved ischemia/reperfu-
6
Department of Anesthesiology,
sion‐induced myocardial injury as evidenced by reduction of the infarct size. Also, during
Anesthesiologist, Faculty of Medicine,
Lorestan University of Medical Science, the ischemic period, ventricular tachycardia and ventricular ectopic beats episodes
Khorramabad, Iran
decreased as compared with that of the control group. The extract stabilized the ST
7
Department of Physiology, School of
Medicine, Gonabad University of Medical segment changes and QTc shortening, decreased R‐wave amplitude, and increased heart
Science, Gonabad, Iran rate during ischemia. The extract also caused significant elevations in serum superoxide
8
Traditional Medicine and History of Medical
dismutase and glutation proxidase activities as well as a significant decrease in serum car-
Sciences Research Center, Institute of Health,
Babol University of Medical Science, Babol, Iran diac troponin I, lactate dehydrogenase, and malondialdehyde levels, 5 days after reperfu-
9
Department of Cardiology, Lorestan sion. In HPLC analysis, the amounts of Cinamic acid, Methyl eugenol, and Cinnamaldehyde
University of Medical Sciences, Khorramabad,
Iran
were 8.99 ± 0.5, 13.02 ± 1.8, and 14.63 ± 1.1 mg/g, respectively. The results show that the
Correspondence ethanolic extract of cinnamon bark is able to protect the heart against ischemia–
Afshin Nazari, Razi Herbal Medicines Research reperfusion injury probably due to its antioxidant properties. Hence, it might be beneficial
Center, Lorestan University of Medical
Science, Khorramabad, Iran. in these patients and this remedy might be used for preparation of new drugs.
Email: nazary257@lums.ac.ir;
nazary257@yahoo.com KEY W ORDS
Funding information
arrhythmia, Cinnamomum zeylanicum, heart, ischemia, rat, reperfusion
Razi Herbal Medicines Research Center,
Lorestan University of Medical Sciences,
Khorramabad, Iran, Grant/Award Number:
Grant

Abbreviations: Catalase, CAT; Cinnamomum zeylanicum, C. zeylanicum; Diastolic arterial pressure, DAP; Glutathione peroxidase, GPX; Heart rate, HR; Infarct sizes/left
ventricles, IS/LV; Ischemia–reperfusion, IR; Lactate dehydrogenase, LDH; Left anterior descending, LAD; Malondialdehyde, MDA; Mean arterial pressure, MAP;
Myocardial infarction, MI; Pulse pressure, PP; Superoxide dismutase, SOD; Systolic arterial pressure, SAP; The duration of action potential, APD; Traditional
Persian medicine, TPM; Triphenyltetrazolium chloride, TTC; Troponin I, cTnI; Ventricular ectopic beat, VEB; Ventricular tachycardia, VT

Phytotherapy Research. 2018;1–9. wileyonlinelibrary.com/journal/ptr Copyright © 2018 John Wiley & Sons, Ltd. 1
2 SEDIGHI
1 | I N T RO D U CT I O N
Myocardial infarction (MI) and its resultant complications are serious
health problems, so common interventions for rapid reperfusion
induction are designed to decrease cardiomyocyte injury and dysfunc-
tion (Nallamothu & Bates, 2003). Although rapid re‐establishment of
cardiac tissue perfusion is an inevitable therapeutic approach, it can
lead to reperfusion injury, which is at least partially related to free rad-
icals (Houshmand, Faghihi, & Zahediasl, 2009). So the use of chemical
or herbal antioxidants has been implicated as a practical approach for
decreasing ischemia–reperfusion (IR) injury in vital organs including
the heart (Ray et al., 1999).
Cinnamon, which botanically refers to Cinnamomum zeylanicum,
is a shrub from family Lauraceae and order Laurales. Cinnamon is
native to Sri Lanka and southern India. It has many uses in food,
medicine, pharmaceutical, and cosmetics industries (Han & Parker,
2017; Madhavadas & Subramanian, 2017; Mesripour, Moghimi, &
Rafieian‐Kopaie, 2016; Nayak, Chinta, & Jetti, 2017; Tulini et al.,
2017; Wong et al., 2016).
Cinnamon has potent antioxidant activity and the chemical
compounds present in cinnamon can prevent the formation of free
radicals (Hassanzadeh, 2012).
There is a growing interest in complementary and alternative
medicine, and experimental studies about the traditionally used drugs
may lead us to new therapeutic approaches in a more cost‐effective
way than random assessments (Fabricant & Farnsworth, 2001;
Rasoulian & Kheirandish, 2017; Wall & Wani, 1996). In present study,
traditional Persian medicine references were assessed, and about 100
herbs were scored based on their therapeutic effects on heart by a
model described previously (Mozaffarpur, Khodadust, Shirafkan,
Yousefi, & Mirzapor, 2016).
Cinnamomum zeylanicum was one of the best materia medica
with the highest scores that could strengthen the heart and improve
its function based on related tradition references. Most of the out-
standing references of traditional Persian medicine include Advieh
Ghalbieh (drugs for heart) of Avicenna, Zakhireh Kharazmshahi (the
store for king of Kharazm) written by Jorjani (Zarshenas, Zargaran,
Abolhassanzadeh, & Vessal, 2012), and Makhzan al‐Advieh (reservoir
of drugs) from Aghili Shirazi (Akhlaghi & Bandy, 2009). Cinnamomum
zeylanicum was selected for this study.
The plant extract and its constituents possess antimicrobial, insec-
ticidal (Dušan, Marián, Katarína, & Dobroslava, 2006; Yang, Lee, Lee,
Clark, & Ahn, 2005), acaricidal (Fichi, Flamini, Zaralli, & Perrucci,
2007), and antimutagenic (Jayaprakasha, Negi, Jena, & Rao, 2007)
activities. In addition, other evidence suggests that cinnamon may be
effective in the treatment of cancer (Nishida et al., 2003) and infec-
tious diseases (Hayashi et al., 2007). It also has anti‐inflammatory
(Tung, Chua, Wang, & Chang, 2008), hypotensive (Preuss, Echard,
Polansky, & Anderson, 2006), and cholesterol‐lowering effects (Khan,
Safdar, Khan, Khattak, & Anderson, 2003). However, little is known
about cardio‐protective actions of C. zeylanicum in and its
antiarithmetic effects.
In experimental arrhythmia, Cinnamomum migao (other species of
cinnamon) reduced the incidence of ventricular fibrillation caused by
chloroform in mice and the ventricular tachycardia (VT) induced
ET AL.
by adrenalin in rabbits, delayed the onset time of this arrhythmia,
increased the arrhythmic doses of strophantin‐K in guinea pigs,
reduced the incidence of some arrhythmias caused by barium chloride
in rats, and slowed down their heart rates (HRs; Sui, Qiu, Xie, & Chen,
1998). According to the traditional use of cinnamon and also its anti-
oxidant property, we assessed the effect of cinnamon bark extract
against ischemia‐induced arrhythmias and heart injury after 5 days
of reperfusion in an in vivo rat model of regional heart ischemia. In this
regard, the effects of cinnamon bark extract were assessed on the
infarct size, VT, and ventricular ectopic beats (VEBs) episodes, the ST
segment changes, QTc shortening, R‐wave amplitude, and HR during
ischemia. The serum superoxide dismutase (SOD) and glutation
proxidase (GPX) activities as well as the serum cardiac troponin I
(cTnI), lactate dehydrogenase (LDH), and malondialdehyde (MDA)
levels were also evaluated. The extract was standardized by measuring
Cinamic acid, Methyl eugenol, and Cinnamaldehyde through HPLC
analysis. The extract antioxidant activity as well as its total flavonoid
and phenolic contents were also determined.
2 | MATERIALS AND METHODS
Healthy Sprague–Dawley male rats weighing 250–300 g were pur-
chased from the Research Center of Lorestan Province. The animals
were maintained in a temperature controlled room at 21–24°C and
40–50% humidity, a 12‐hr light/dark cycle, and free access to food
and water.
Cinnamomum zeylanicum samples were purchased from local gro-
ceries in Shahrekord city, southwest Iran in April–May and authenti-
cated as C. zeylanicum by a botanist. All chemical compositions were
obtained from Sigma‐Aldrich (USA) unless otherwise stated.
2.1 | Preparation of plant material
Coarse powder from dried barks of C. zeylanicum was extracted to
exhaustion with ethanol (96%) using a Soxhlet apparatus with ethanol
solvent systems. The extract was dissolved in sterilized distilled water
before oral administration to the experimental animals.
Chemical analysis of C. zeylanicum was determined by HPLC
method consisted of a quaternary pump (LC‐10ATvp), manual injector
with a 10‐μl sample loop, and a C18 analytical column (Shimadzu,
150 × 4.6 mm id, 5‐μm particle size; Japan). Effluent was monitored
by UV‐Vis detector (SPD‐M10Avp) at 280 nm. The mobile phase
consisted of water (containing 0.1% v/v of acetic acid) and acetonitrile
(80/20, v/v). The mobile phase components, including water and ace-
tonitrile, were degassed separately by a Millipore vacuum pump prior
usage. Flow rate and column oven were set at 1.0 ml/min and ambient
temperature, respectively.
2.2 | Determination of total phenolic content
The total phenolic content of the hydroethanolic extract of
C. zeylanicum was determined using the Foline–Ciocalteu method as
modified by (Singleton, Orthofer, & Lamuela‐Raventós, 1999).
SEDIGHI ET AL.
2.3 | Determination of total flavonoid content
The total flavonoid content was determined using a colorimetric
method described by Dewanto (Chang, 2002).
2.4 | Assay for total antioxidant activity
Reduction of the stable free radical 2,2‐diphenyl‐1‐picrylhydrazyl
(DPPH) was determined with the aid of a modified version of the
method described by Shimada, Fujikawa, Yahara, and Nakamura
(1992). The result was expressed as a percentage of the absorbance
of a control DPPH solution without test compounds.
2.5 | Surgical preparation
Anesthesia was achieved in animals by administration of sodium thio-
pental (60‐mg/kg body weight, i.p.). The animals were placed on
operating table, and their chests were completely shaved. During the
surgery, the temperature of the rats' bodies was maintained at
37 ± 1°C using a thermoregulator. The rats' necks were positioned in
a way that tracheal intubation could be easily conducted on them.
After intubation, the animals were ventilated at 60–70 breaths/min
and tidal volume 1.5 cc/kg. To record electrocardiography (ECG) lead
II, subcutaneous needle electrode was used such that the negative
electrode was subcutaneously connected to the right arm, the positive
electrode to the left foot, and the neutral electrode to the left arm.
Then an incision was made on the left fourth intercostal space of
the chest so carefully that the left lung and heart were not injured.
FIGURE 1 Examples of electrocardiogram recordings and definition of various changes and arrhythmias: (a) During baseline; (b) ST elevation
during coronary artery occlusion; (c) ventricular ectopic beat; (d) couplet; (e) bigeminy; (f) ventricular tachycardia (VT); (g) ventricular fibrillation
(VF); (h) ECG parameters [Colour figure can be viewed at wileyonlinelibrary.com]
3
Afterwards, pericardium was slowly torn, and silk thread 0.6 carefully
passed through under left anterior descending and fastened with
ligature apparatus (Bavanesh: a device useful for coronary ligation in
rats, Elmbavaran Aftab Lorestan Company, Lorestan, Iran).
Reperfusion was induced by dragging and releasing the thread of
Bavanesh. After completion of the ischemia and beginning of the
reperfusion, the incision was closed, tracheal tube removed, and
the rats completely oxygenated and recovered. Two criteria, that is,
disorder in normal myocardial contraction and changes in ST segment
in ECG, were considered to confirm induction of MI. After complete
recovery, the rats were caged, fed with food and water, and
transferred to Animal House. Except for arrhythmias, which were
examined during ischemic period, other variables were examined at
the end of the fifth day after reperfusion. Tetracycline ointment
was used as antibiotic.
2.6 | Experimental protocol
This study was approved by the laboratory animal center of the Razi
Herbal Medicines Research Center, Lorestan University of Medical
Sciences, Khorramabad (ethical code: LUMS.REC.1394.79).
The rats were randomly assigned to four groups (eight rats in
each). Three groups were given C. zeylanicum (CZ) extract in three
different doses of 50, 100, and 200 mg·kg·day. A control group was
given distilled water. All treatments were oral. The rats were
pretreated for 14 days and then underwent a 30‐min regional heart
4 SEDIGHI ET AL.

ischemia and a 5‐day reperfusion period. Blood samples were col- The QT interval was measured starting from the onset of the QRS
lected by cardiac puncture for serum enzyme assays. complex until the end of the T wave. QTc was obtained using Bazett's
formula (QTc = QT/√RR; Bazett, 1920).

2.7 | Hemodynamic functions

The rats were anesthetized again 5 days after reperfusion by keta- 2.10 | Determination of MDA, LDH, cTnI, CAT, SOD,
mine–xylazine (100 mg/kg and 10 mg/kg, respectively) administration, and GPX in the serum
and then the hemodynamic parameters such as systolic arterial
Blood samples were collected at the end of fifth day of the reperfu-
pressure (SAP), diastolic arterial pressure (DAP), HR, calculated
sion for measurement of LDH (as a heart injury marker), SOD, GPX,
pulse pressure (i.e., the difference of SAP and DAP), and mean arterial
and catalase (CAT; as three antioxidant enzymes) activities and MDA
pressure were recorded.
(as a lipid peroxidation marker) and cTnI (as a specific cardiac injury
biomarker) levels.
2.8 | Determination of infarct size The heparinized samples were centrifuged at 5,000 rpm, for
15 min, and the plasma was removed and stored at −70°C until the
At the end of experiment, the heart was excised, and both atria
time that they were assayed. The activity of LDH and the cTnI level
and the roots of the great vessels were removed. The heart was
were analyzed using commercial kits (Pars Azmoon, Iran, Zellbio) by
frozen overnight and then cut into slices of 2 mm thick. All slices were
employing an autoanalyzer (Roche Hitachi Modular DP Systems,
incubated with a 1% solution of 2,3,5‐triphenyltetrazolium chloride (in
Mannheim, Germany). MDA content of samples was determined
0.1‐M phosphate buffer, pH 7.4) and stained for 15 min at 37°C to
spectrophotometrically using a modification of the assay described
visualize the infarct area. Then they were fixed in 10% formalin. Both
by (Schuh, Fairclough, & Haschemeyer, 1978). SOD, CAT, and GPX
surfaces of each section were scanned using PhotoShop program
activities were measured spectrophotometrically using diagnostic kits
(Adobe Systems, version 7.0). Infarct size percent was expressed as a
according to the manufacturers' instructions (Chance, 1995).
percentage of the left ventricles (IS/LV).

2.9 | Determination of arrhythmias and 2.11 | Statistical analysis

electrocardiogram variability
All of the data are presented as the mean ± SEM. Differences among
During the 30‐min ischemia, ventricular arrhythmias were evaluated. groups were determined by one‐way analysis of variance, and
VEBs were defined as identifiable premature QRS complexes. Ventric- p < 0.05 was considered a statistically significant.
ular tachycardia was defined as a run of four or more ventricular pre-
mature beats, and the ventricular fibrillation was defined as a signal for
which individual QRS deflections could no longer be distinguished 3 | RESULTS
from each other and for which a rate could no longer be measured.
Original ECG recordings are illustrated in Figure 1. The incidence,
3.1 | Radical scavenging activity, total flavonoids,
onset time, and duration of arrhythmias were assigned to identify
and total phenolic content of C. zeylanicum bark
arrhythmias severity according to the following scoring system (Curtis
& Walker, 1988) The average QRS interval (Figure 1h) duration,
extract
corrected QT interval (QTc) time, T and R amplitudes, and ST‐segment The antioxidant activity of the extract was 61.31% of the DPPH, and
elevations were recorded at baseline, at the end of 30‐min ischemia the flavonoids and total phenolic compounds were respectively 8.32
and 60 min of reperfusion. The QRS complexes were analyzed. and 27.9 mg/g of dried extract.

FIGURE 2 HPLC chromatographic pattern

for simultaneous determination of
Cinnamaledhyde, Cinamic acid, and Methyl
eugenol from extract of Cinamomum
zeylanicum bark powder [Colour figure can be
viewed at wileyonlinelibrary.com]
SEDIGHI ET AL. 5

TABLE 1 Effect of Cinamomum zeylanicum (CZ) bark on heart rate

(beats/min) in various groups

Groups Baseline End of ischemia 30' End of reperfusion 60'

Control 205.87 ± 15.97 220.5 ± 29.41 179.62 ± 22.38
CZ‐50 155.75 ± 16.40 241.62 ± 15.95** 196.5 ± 17.45
CZ‐ 165.25 ± 27.10 226.37 ± 14**£ 219.5 ± 21.21
100
CZ‐ 145.87 ± 13.87 314.4 ± 15.80**# 192.25 ± 27.58$$
200

Note. The values are as mean ± SEM (n = 8 in all groups).

**Significant difference compared to its baseline (p < 0.01);
#
Significant intergroup differences compared with control group (p < 0.05); FIGURE 3 The original pictures of TTC staining of heart slices.
$$
Significant difference compared with the end of ischemic period Myocardial infarct size (IS/LV%) in control and CZ (50, 100, and 200)
(p < 0.01); groups. Data are presented as mean ± SEM. ***, p < 0.001 versus
£
Significant intergroup differences compared with CZ‐200 group control and CZ‐50 groups. ### p < 0.001 versus CZ (100 and 200)
(p < 0.05). groups. CZ: Cinnamomum zeylanicum extract‐treated groups [Colour
figure can be viewed at wileyonlinelibrary.com]

3.2 | HPLC analysis of C. zeylanicum 3.4 | Infarct size measurements

The results of HPLC analysis demonstrated that the major constitu- Infarct size was 43.22 ± 20% in control group. Administration of
ents of the bark extract consisted of Cinamic acid (8.99 ± 0.5 mg/g), higher doses of CZ (100 and 200 mg/kg) significantly reduced infarct
Methyl eugenol (13.02 ± 1.8 mg/g), and Cinnamaldehyde sizes (32.89 ± 0.92% and 28.03 ± 0.83%, respectively) compared with
(14.63 ± 1.1 mg/g), respectively. Figure 2 shows the chromatogram
of the chemical compounds of the sample.

3.3 | Hemodynamic functions

Table 1 demonstrates the HR during the experiments at specific times.
There were no significant differences among groups at baseline
(before ischemia).
At the end of 30‐min ischemia, the HR significantly increased in
treatment (50, 100, and 200) groups compared with their baseline
values and was significantly higher in CZ‐200 group compared with
the control and CZ‐100 groups. In CZ‐200 group, HR was significantly
decreased at the end of 60‐min reperfusion compared with its value at
the end of ischemic period.
Table 2 demonstrates the time course of mean arterial pressure,
SAP, DAP, and pulse pressure 5 days after reperfusion. DAP
significantly decreased in treatment CZ‐50 group compared with the
control group. There were no significant differences between any
other groups.

TABLE 2 Effect of Cinamomum zeylanicum (CZ) bark on intravas-

cular hemodynamic parameters 5 days after reperfusion in different
groups
MAP SAP DAP PP
Groups (Hg/mm) (Hg/mm) (Hg/mm) (Hg/mm)
Control 87 ± 6.88 106.5 ± 5.94 73.87 ± 7.25 42.75 ± 1.25
FIGURE 4 Ventricular arrhythmias during ischemia. (a) The total
CZ‐50 81.62 ± 10.35 99 ± 7.09 52.5 ± 4.02* 42.62 ± 7.26
number of ventricular tachycardia (VT) and ventricular ectopic beats
CZ‐100 72.12 ± 7.24 82 ± 4.63 71.37 ± 2.79 25.75 ± 3.43
(VEBs) during 30‐min ischemia in different groups. (b) Duration (s) of
CZ‐200 72.33 ± 7.31 104.66 ± 15.16 72.33 ± .8.66 20.66 ± 3.48 VT during 30‐min ischemia in different groups. Data are presented as
*Significant difference compared with the control group (p < 0.05). The mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 versus control group,
values are as mean ± SEM. MAP, SAP, and DAP (mean, systolic, and dia- #p < 0.05 versus CZ (50 and 100) groups. CZ: Cinnamomum zeylanicum
stolic arterial pressures, respectively) and PP (pulse pressure; n = 8 in all extract‐treated groups [Colour figure can be viewed at
groups). wileyonlinelibrary.com]
6 SEDIGHI ET AL.

control and CZ‐50 (42.25 ± 0.72%) groups. Lower dose of CZ (50 mg/
kg) had no effect on infarct size (Figure 3).
3.5 | Ventricular arrhythmias during ischemia
Figure 4a shows the total number of VEBs and VTs. The mean number
of VEB episodes during 30 min of ischemia in CZ‐100 and CZ‐200
groups (41.25 ± 11.3 and 30.12 ± 6.6, respectively) markedly reduced
compared with control and CZ‐50 groups (72.37 ± 8.03 and
50.75 ± 11.14, respectively). Treatment with Cinamomum zeylanicum
extract in CZ‐200 group reduced the number of VT episodes, com-
pared with control.
Figure 4b shows the total duration of VT episodes. Compared
with control rats, total duration of VTs throughout 30 min of ischemia
was significantly reduced by administration of C. zeylanicum extract in
CZ‐200 group (65.62 ± 38.1 s vs. 369.5 ± 49.9 s). In CZ‐50 and CZ‐
100 groups, there was not any significant difference compared with
control group in this regard.
3.6 | ECG parameters
The ECG analysis showed noticeable variations including mean QTc
interval shortening in treatment CZ (50 and 100) groups at the end
of ischemic period and 60 min of reperfusion compared with their
baseline values. There was conversely prolongation of QTc in CZ‐
200 at the end of ischemic period compared with its baseline. QRS
interval was shorted in treatment CZ‐100 group at the end of ischemia
period compared with its baseline. Repolarization voltage (T‐wave
amplitude) was significantly increased in treatment CZ‐200 group at
end of ischemia period compared with its baseline (Table 3).
Intergroup comparisons showed that at baseline, duration of QTc
was significantly lower in CZ‐200 group as compared with control
group. Also QRS interval was significantly shorter in CZ‐200 group
as compared with CZ‐50 group.
At the end of ischemia, QTc duration and QRS amplitude signif-
icantly decreased in treatment CZ (50,100, or 200) groups as
compared with control group. Also, R‐wave amplitude significantly
decreased in CZ‐200 group as compared with control group and
T‐wave amplitude significantly increased in CZ‐200 group as com-
pared with control and CZ‐50 groups. Administration of C. zeylanicum
extract in CZ (100 or 200) groups markedly stabilized ST segment
changes, too.
At end of reperfusion period, QTc significantly decreased in
treatment CZ (50, 100, and 200) groups as compared with control
group and QRS amplitude significantly decreased in treatment
CZ‐200 group as compared with control group. Also R‐wave ampli-
tude significantly decreased in CZ (50, 100, or 200) groups compared
with control group. In this period, T‐wave amplitude significantly
increased in CZ‐200 group compared with CZ‐50 group and ST
segment elevation decreased in CZ‐200 compared with control
group (Table 3).
3.7 | Biochemical analysis
3.7.1 | Antioxidant enzyme activities
Catalase, SOD, and GPX activities 5 days after reperfusion were exam-
ined in serum samples in various experimental groups. As shown in
Table 4, SOD and GPX activities significantly increased in the CZ
(100 and 200) groups compared with CZ‐50 and control groups
(p < 0.05).

TABLE 3 Effect of Cinamomum zeylanicum (CZ) bark on ECG parameters in various groups

Time Variable Control CZ‐50 CZ‐100 CZ‐200

Baseline QTC (ms) 242.25 ± 24.86 221.25 ± 15.33 211.37 ± 16.5 104.12 ± 2.7###$$$
QRS (μV) 17.37 ± 0.65 19 ± 1.25 17.75 ± 0.67 15.62 ± 0.46$
R (μV) 504.12 ± 47.56 463.75 ± 59.15 415.62 ± 80.27 383.42 ± 95.60
T (μV) 267.87 ± 48.16 210.62 ± 56.13 224 ± 23.29 241.37 ± 23.75
ST (μV) 275 ± 20.08 221.25 ± 32.38 182.12 ± 32.38 154.37 ± 27.49#
End of ischemia 30 QTC (ms) 279.75 ± 21.89 121.37 ± 12.1***### 109.62 ± 10.65***### 162.12 ± 29.60**###
QRS (μV) 21.87 ± 2.03 17 ± 0.65# 12.25 ± 0.64**### 14 ± 0.65##
R (μV) 630.37 ± 98.94 419.25 ± 64.78 402 ± 37.79 209.5 ± 30.13###
T (μV) 245.52 ± 19.05 256.25 ± 13.06£ 265.75 ± 25.75 364.5 ± 38.42*#
ST (μV) 319.75 ± 44.73 249.37 ± 18.1 209 ± 22.03## 173.12 ± 20.02##
End of reperfusion 60 QTC (ms) 250.62 ± 20.07 136.62 ± 12.74***### 131 ± 9.04***### 106.12 ± 4.47###
QRS (μV) 19.62 ± 1.22 17.12 ± 1.38 16.37 ± 0.65 14.5 ± 0.42##
R (μV) 542.87 ± 44.52 383.62 ± 22.5# 354.25 ± 15.71## 254.87 ± 52###
T (μV) 250.75 ± 22.51 209.52 ± 14.4£ 264.5 ± 38.428 357 ± 36.70#
ST (μV) 314 ± 29.42 214.75 ± 42.19 188 ± 31.89 118.75 ± 28.74##

*p < 0.05;
**p < 0.01; and
***p < 0.001 significant difference compared with its baseline.
#
p < 0.05;
##
p < 0.01; and
###
p < 0.001 significant intergroup differences compared with control group.
$
Significant intergroup differences compared with CZ‐50 group (p < 0.05).
$$$
Significant intergroup differences compared with CZ‐50 and CZ‐100 groups (p < 0.001).
£
Significant inter group differences compared with CZ‐200 group (p < 0.05). The values are mean ± SEM.
SEDIGHI ET AL. 7

TABLE 4 Effect of Cinamomum zeylanicum (CZ) bark on serum cat- coronary heart disease (Hertog, Feskens, Kromhout, Hollman, &
alase (CAT), superoxide dismutase (SOD), and glutathione peroxidase Katan, 1993). The most important property of the flavonoids is the
(GPX), 5 days after reperfusion antioxidant activity that could be due to scavenging of free radicals,
Groups CAT (unit/mg) SOD (unit/mg) GPX (unit/mg) interfering with inducible nitric‐oxide synthase activity and inhibition
Control 105.26 ± 3.24 56.62 ± 2.66 686.17 ± 142.5 of xanthin‐oxidase (Nijveldt et al., 2001). So the antioxidant
CZ‐50 135.5 ± 9.03 59.37 ± 5.78 782.27 ± 148.7 properties and flavonoid contents of the extract may have important
CZ‐100 147.3 5 ± 6.95 99.71 ± 19.45* #
811.97 ± 53.18* # roles in preventing I/R‐induced injuries such as arrhythmia and
CZ‐200 186.37 ± 3558 110.83 ± 22.67* #
826.25 ± 53.92* # infarction (Najafi, Ghasemian, Fathiazad, & Garjani, 2009). Previous
studies have also shown that flavonoids in Cinnamomum extract
Note. The values are mean ± SEM of eight animals in each group.
may be responsible for its antioxidant and thus, for example, its
*Significant difference compared with control (p < 0.05);
#
hepatoprotective activity through inhibition of lipid peroxidation
Significant difference compared with CZ‐50 group (p < 0.05, (n = 8 in all
groups). against CCL4‐induced liver injury in rats (Eidi, Mortazavi, Bazargan,
& Zaringhalam, 2012).
3.7.2 | CTnI level and LDH activity and lipid peroxi- The major compounds of the bark of C. zeylanicum sample were
dation level determined by HPLC. A major polyphenolic component of the plant
Figure 5a,b shows that administration of C. zeylanicum extract in CZ sample was Methyl eugenol. Methyl eugenol has certain pharmacolog-
(100 and 200) groups prevents elevation of cTnI level and dehydroge- ical properties including anesthetic activity, antioxidant potential,
nase (LDH) activity in serum, 5 days after ischemia/reperfusion. Also,
serum MDA level in CZ (50, 100, and 200) groups was significantly
declined compared with control group (Figure 5c)

4 | DISCUSSION

According to the present study results, the ethanolic C. zeylanicum

bark extract significantly decreased IR‐induced myocardial injury as
evidenced by reducing the infarct size. Also the episodes of ische-
mia‐induced VTs and VEBs and total duration of VTs were reduced
compared with those of the control group. It also stabilized the ST
segment changes and QTc shortening, decreased the R‐wave ampli-
tude, increased heart rate during ischemia, increased serum SOD and
GPX activities, and decreased serum cTnI, LDH, and MDA levels 5 days
after reperfusion.
Since more than two decades ago, the role of reactive oxygen
species in many cardiovascular diseases has become increasingly
apparent. Under normal conditions, there is a balance between the
formation of oxygen‐free radicals and the amount of antioxidants.
This steady‐state condition may be interrupted in some pathophysi-
ologic conditions, like an ischemic insult and subsequent reperfusion,
because of the excessive production of free radicals and/or decrease
in antioxidants (Arpad, Bagchi, Pali, Cordis, & Das, 1993; Lowenstein,
2007). A significant amount of evidence is present in the literature
to support the role of oxygen free radicals in pathogenesis of myo-
cardial ischemia reperfusion injury, too (Das et al., 1986). This
receives further support from this fact that many free radical
scavengers and antioxidants are capable of ameliorating IR injury
(Das & Maulik, 1994). Total phenolic compounds and flavonoid
content of dried Cinnamomum bark extract used in present study
showed relatively high amounts of these compounds and high level
of antioxidant activity. Therefore, the cardioprotective effects of
effects of C. zeylanicum, at least in part, might be attributed to its FIGURE 5 Reperfusion in different groups. The values are expressed
as mean ± SEM; cTnI: cardiac troponin I; LDH: lactate dehydrogenase;
antioxidant activity.
MDA: malondialdehyde. Data are presented as mean ± SEM.*
There is an inverse association between flavonoid intakes Significant difference compared with control (p < 0.05), ** Significant
and coronary heart disease mortality (Nijveldt et al., 2001). Also, a difference compared with control (p < 0.01). [Colour figure can be
clinical study showed that flavonoids may reduce mortality from viewed at wileyonlinelibrary.com]
8 SEDIGHI
antimicrobial role, anti‐inflammatory action, anticarcinogenic effects,
neuroprotective ability, hypolipidemic, and antidiabetic effects (Khalil
et al., 2017).
Phenolic compounds have hydroxyl groups on their aromatic ring
(s) and exhibit antioxidant, antimutagenic, and carcinogenic activities,
which have been attributed to their scavenger activities against ROS
(Rice‐Evans, Miller, & Paganga, 1996).
As it was said, the protective effects of C. zeylanicum bark extract
in present study may be related at least in part to the above‐men-
tioned antioxidant effect of this herbal extract. This could be sup-
ported by the higher level of the serum antioxidant enzymes, SOD
and GPX activities, and lower level of MDA following administration
of Cinnamomum extract compared with control group. It should be
noted that serum MDA level has been used as an indicator of lipid per-
oxidation and tissue damage caused by in vivo free oxygen radicals
(Kim, Kwack, & Lee, 2000.
Ventricular arrhythmias are the most important causes of mortal-
ity after cardiac surgery and MI (Najafi, Ghasemian, Fathiazad, &
Garjani, 2009). In addition to its culinary uses, cinnamon has been
employed in traditional herbal medicine to treat a variety of health
conditions including heart failure (Zarshenas, Zargaran,
Abolhassanzadeh, & Vessal, 2012). IKr channels play a role in cardiac
excitability and usually are selected as a target for detecting the effect
of antiarrhythmic compounds. However, one of the major drawbacks
of these agents is the prolongation of QT interval, which can result
in lethal arrhythmias, such as torsade de pointes. This effect makes
these drugs potentially dangerous (Dong et al., 2004). So developing
antiarithmetic drugs devoid of the QT prolongation is one of the main
targets for many researchers. As was mentioned, the antiarithmetic
property of C. zeylanicum extract in present study was associated with
a significant decrease of QT interval.
Administration of C. zeylanicum caused a decrease in R‐wave
amplitude, which represents the cardiac contractility (Pinsky et al.,
1995). So it can be concluded that C. zeylanicum extract may be a neg-
ative inotropic agent due to decrease in myocardial contractility.
Administration of C. zeylanicum significantly reduced the levels of
injury markers including cTnI and LDH. Especially, cTnI, an excellent
serum marker for detecting myocardial injury, was significantly
reduced by C. zeylanicum extract administration, supporting our data
about lower infarct size in this group.
5 | C O N CL U S I O N
The ethanolic bark extract of C. zeylanicum exhibited cardio‐protec-
tive effects against ischemia‐induced arrhythmias and cardiac injury
as was reflected by a reduction in infarct size and cardiac injury
biomarkers. The antioxidant properties of cinnamon extracts, which
are partially attributable to the ability of its phenolic constituents
to quench reactive oxygen species, are thought to contribute in
these protective properties. These data support the potential uses
of cinnamon in the treatment of heart IR disorders. Hence, it
might be used for development of new antiarithmetic drugs after
further investigations.
ET AL.
ACKNOWLEDGMENTS
This study was supported financially by Razi Herbal Medicines
Research Center, Lorestan University of Medical Sciences, Khorrama-
bad, Iran.
CONFLIC T OF INT E RE ST
The authors declare no conflict of interest.
ORCID
Mehrnoosh Sedighi http://orcid.org/0000-0002-2002-2413
RE FE RE NC ES
Akhlaghi, M., & Bandy, B. (2009). Mechanisms of flavonoid protection
against myocardial ischemia–reperfusion injury. Journal of Molecular
and Cellular Cardiology, 46(3), 309–317.
Arpad, T., Bagchi, D., Pali, T., Cordis, G. A., & Das, D. K. (1993). Compari-
sons of ESR and HPLC methods for the detection of OH. radicals in
ischemic/reperfused hearts: A relationship between the genesis of free
radicals and reperfusion arrhythmias. Biochemical Pharmacology, 45(4),
961–969.
Bazett, H. C. (1920). An analysis of the time relations of electrocardio-
grams. Heart, 7, 353–370.
Chance, C. M. (1995). Assay of catalase and peroxidases. Methods of
Enzymology, 11, 764–775.
Chang, H.‐J. (2002). Kicking away the ladder: Development strategy in
historical perspective. ( p. 196) London: Anthem Press.
Curtis, M., & Walker, M. J. (1988). Quantification of arrhythmias using
scoring systems: An examination of seven scores in an in vivo model
of regional myocardial ischaemia. Cardiovascular Research, 22,
656–665.
Das, D., Engelman, R., Rousou, J., Breyer, R., Otani, H., & Lemeshow, S.
(1986). Pathophysiology of superoxide radical as potential mediator
of reperfusion injury in pig heart. Basic Research in Cardiology, 81(2),
155–166.
Das, DK, Maulik, N. 1994. [60] Antioxidant effectiveness in ischemia‐
reperfusion tissue injury. In: Methods in Enzymology, Elsevier; pp.
601–610.
Dong, D. L., Li, Z., Wang, H. Z., Du, Z. M., Song, W. H., & Yang, B. F. (2004).
Acidification alters antiarrhythmic drug blockade of the ether‐a‐go‐go‐
related gene (HERG) channels. Basic & Clinical Pharmacology & Toxicol-
ogy, 94(5), 209–212.
Dušan, F., Marián, S., Katarína, D., & Dobroslava, B. (2006). Essential oils
—Their antimicrobial activity against Escherichia coli and effect on
intestinal cell viability. Toxicology In Vitro, 20(8), 1435–1445.
Eidi, A., Mortazavi, P., Bazargan, M., & Zaringhalam, J. (2012). Hepatopro-
tective activity of cinnamon ethanolic extract against CCI4‐induced
liver injury in rats. EXCLI Journal, 11, 495–507.
Fabricant, D. S., & Farnsworth, N. R. (2001). The value of plants used in tra-
ditional medicine for drug discovery. Environmental Health Perspectives,
109(Suppl 1), 69–75.
Fichi, G., Flamini, G., Zaralli, L., & Perrucci, S. (2007). Efficacy of an
essentifal oil of Cinnamomum zeylanicum against Psoroptes cuniculi.
Phytomedicine, 14(2), 227–231.
Han, X., & Parker, T. L. (2017). Antiinflammatory activity of cinnamon
(Cinnamomum zeylanicum) bark essential oil in a human skin disease
model. Phytotherapy Research, 31, 1034–1038.
Hassanzadeh, A. (2012). Cinnamon and its properties. Monthly training and
new technology. Research of Food, (22), 37–39.
Hayashi, K., Imanishi, N., Kashiwayama, Y., Kawano, A., Terasawa, K.,
Shimada, Y., & Ochiai, H. (2007). Inhibitory effect of cinnamaldehyde,
derived from Cinnamomi cortex, on the growth of influenza A/PR/8
virus in vitro and in vivo. Antiviral Research, 74(1), 1–8.
SEDIGHI ET AL.
Hertog, M. G., Feskens, E. J., Kromhout, D., Hollman, P., & Katan, M.
(1993). Dietary antioxidant flavonoids and risk of coronary heart dis-
ease: The Zutphen Elderly Study. The Lancet, 342(8878), 1007–1011.
Houshmand, F., Faghihi, M., & Zahediasl, S. (2009). Biphasic protective
effect of oxytocin on cardiac ischemia/reperfusion injury in anaesthe-
tized rats. Peptides, 30(12), 2301–2308.
Jayaprakasha, G., Negi, P., Jena, B., & Rao, L. J. M. (2007). Antioxidant and
antimutagenic activities of Cinnamomum zeylanicum fruit extracts.
Journal of Food Composition and Analysis, 20(3), 330–336.
Khalil, A. A., Ur Rahman, U., Khan, M. R., Sahar, A., Mehmood, T., & Khan,
M. (2017). Essential oil eugenol: Sources, extraction techniques and
nutraceutical perspectives. RSC Advances, 7(52), 32669–32681.
Khan, A., Safdar, M., Khan, M. M. A., Khattak, K. N., & Anderson, R. A.
(2003). Cinnamon improves glucose and lipids of people with type 2
diabetes. Diabetes Care, 26(12), 3215–3218.
Kim, H. S., Kwack, S. J., & Lee, B. M. (2000). Lipid peroxidation, antioxidant
enzymes, and benzo [a] pyrene‐quinones in the blood of rats treated
with benzo [a] pyrene. Chemico‐Biological Interactions, 127(2),
139–150.
Lowenstein, C. (2007). Myocardial reperfusion injury. The New England
Journal of Medicine, 357, 2409–2410.
Madhavadas, S., & Subramanian, S. (2017). Cognition enhancing effect of
the aqueous extract of Cinnamomum zeylanicum on non‐transgenic
Alzheimer's disease rat model: Biochemical, histological, and behav-
ioural studies. Nutritional Neuroscience, 20(9), 526–537.
Mesripour, A., Moghimi, F., & Rafieian‐Kopaie, M. (2016). The effect of
Cinnamomum zeylanicum bark water extract on memory performance
in alloxan‐induced diabetic mice. Research in Pharmaceutical Sciences,
11(4), 318–323.
Mozaffarpur, S. A., Khodadust, M., Shirafkan, H., Yousefi, M., & Mirzapor,
M. (2016). Introducing a model for prioritization of drugs, based on
Iranian traditional medicine references. History of Medicine Journal
(Quarterly), 6(19), 11–28.
Najafi, M., Ghasemian, E., Fathiazad, F., & Garjani, A. (2009). Effects of
total extract of Dracocephalum moldavica on ischemia/reperfusion
induced arrhythmias and infarct size in the isolated rat heart. Iranian
Journal of Basic Medical Sciences, 11(4), 229–235.
Nallamothu, B. K., & Bates, E. R. (2003). Percutaneous coronary interven-
tion versus fibrinolytic therapy in acute myocardial infarction: Is
timing (almost) everything? The American Journal of Cardiology, 92(7),
824–826.
Nayak, I. N., Chinta, R., & Jetti, R. (2017). Anti‐atherosclerotic potential of
aqueous extract of cinnamomum zeylanicum bark against glucocorti-
coid induced atherosclerosis in wistar rats. Journal of Clinical and
Diagnostic Research: JCDR, 11(5), FC19–FC23.
Nijveldt, R. J., Van Nood, E., Van Hoorn, D. E., Boelens, P. G., Van Norren,
K., & Van Leeuwen, P. A. (2001). Flavonoids: A review of probable
mechanisms of action and potential applications. The American Journal
of Clinical Nutrition, 74(4), 418–425.
Nishida, S., Kikuichi, S., Yoshioka, S., Tsubaki, M., Fujii, Y., Matsuda, H., …
Irimajiri, K. (2003). Induction of apoptosis in HL‐60 cells treated with
medicinal herbs. The American Journal of Chinese Medicine, 31(04),
551–562.
Pinsky, M. R., Gorcsan, J., Gasior, T. A., Mandarino, W. A., Deneault, L. G.,
Hattler, B. G., & Kunig, H. (1995). Changes in electrocardiographic mor-
phology reflect instantaneous changes in left ventricular volume and
output in cardiac surgery patients. The American Journal of Cardiology,
76(10), 667–674.
Preuss, H. G., Echard, B., Polansky, M. M., & Anderson, R. (2006). Whole
cinnamon and aqueous extracts ameliorate sucrose‐induced blood
9
pressure elevations in spontaneously hypertensive rats. Journal of the
American College of Nutrition, 25(2), 144–150.
Rasoulian, B., & Kheirandish, F. (2017). Herbal medicines: From traditional
medicine to modern experimental approaches. Herbal Medicines
Journal, 2(1), 1–2.
Ray, P. S., Maulik, G., Cordis, G. A., Bertelli, A. A., Bertelli, A., & Das, D. K.
(1999). The red wine antioxidant resveratrol protects isolated rat
hearts from ischemia reperfusion injury. Free Radical Biology and
Medicine, 27(1), 160–169.
Rice‐Evans, C. A., Miller, N. J., & Paganga, G. (1996). Structure‐antioxidant
activity relationships of flavonoids and phenolic acids. Free Radical
Biology and Medicine, 20(7), 933–956.
Schuh, J., Fairclough, G. F., & Haschemeyer, R. H. (1978). Oxygen‐medi-
ated heterogeneity of apo‐low‐density lipoprotein. Proceedings of the
National Academy of Sciences, 75(7), 3173–3177.
Shimada, K., Fujikawa, K., Yahara, K., & Nakamura, T. (1992). Antioxidative
properties of xanthan on the autoxidation of soybean oil in cyclodex-
trin emulsion. Journal of Agricultural and Food Chemistry, 40(6),
945–948.
Singleton, VL, Orthofer, R, Lamuela‐Raventós, RM. 1999. [14] Analysis of
total phenols and other oxidation substrates and antioxidants by means
of folin‐ciocalteu reagent. In: Methods in Enzymology, Elsevier 299:
152–178.
Sui, Y., Qiu, D., Xie, C., & Chen, K. (1998). Observation of antiarrhythmic
effects of Cinnamomum migao HW Li on experimental arrhythmia.
Zhongguo Zhong yao za zhi= Zhongguo zhongyao zazhi= China Journal
of Chinese Materia Medica, 23(8), 495–497.
Tulini, F. L., Souza, V. B., Thomazini, M., Silva, M. P., Massarioli, A. P.,
Alencar, S. M., … Favaro‐Trindade, C. S. (2017). Evaluation of the
release profile, stability and antioxidant activity of a
proanthocyanidin‐rich cinnamon (Cinnamomum zeylanicum) extract
co‐encapsulated with α‐tocopherol by spray chilling. Food Research
International, 95, 117–124.
Tung, Y.‐T., Chua, M.‐T., Wang, S.‐Y., & Chang, S.‐T. (2008). Anti‐inflamma-
tion activities of essential oil and its constituents from indigenous
cinnamon (Cinnamomum osmophloeum) twigs. Bioresource Technology,
99(9), 3908–3913.
Wall, M. E., & Wani, M. C. (1996). Camptothecin and taxol: From discovery
to clinic. Journal of Ethnopharmacology, 51(1–3), 239–254.
Wong, H. Y., Tsai, K., Liu, Y. H., Yang, S., Chen, T. W., Cherng, J., … Cherng,
J. M. (2016). Cinnamomum verum component 2‐
methoxycinnamaldehyde: A novel anticancer agent with both anti‐
topoisomerase I and II activities in human lung adenocarcinoma a549
cells in vitro and in vivo. Phytotherapy Research, 30(2), 331–340.
Yang, Y.‐C., Lee, H.‐S., Lee, S. H., Clark, J. M., & Ahn, Y.‐J. (2005). Ovicidal
and adulticidal activities of Cinnamomum zeylanicum bark essential oil
compounds and related compounds against Pediculus humanus capitis
(Anoplura: Pediculicidae). International Journal for Parasitology, 35(14),
1595–1600.
Zarshenas, M. M., Zargaran, A., Abolhassanzadeh, Z., & Vessal, K. (2012).
Jorjani (1042–1137). Journal of Neurology, 1‐2, 6637–6639.
How to cite this article: Sedighi M, Nazari A, Faghihi M, et al.
Protective effects of cinnamon bark extract against ischemia–
reperfusion injury and arrhythmias in rat. Phytotherapy
Research. 2018;1–9. https://doi.org/10.1002/ptr.6127