Cardioprotective Effect of Dichloromethane Valerian (Valeriana Officinalis) Extract on Ischemia-Reperfusion-Induced Cardiac Injuries in Rats

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GENERAL ENDOCRINOLOGY
doi: 10.4183/aeb.2023.178
Acta Endocrinologica (Buc), vol. XVIV, no. 2, p. 178-186, 2023
178
CARDIOPROTECTIVE EFFECT OF DICHLOROMETHANE VALERIAN (VALERIANA
OFFICINALIS) EXTRACT ON ISCHEMIA-REPERFUSION-INDUCED CARDIAC INJURIES IN RATS
M. Sedighi1, H. Seidi2, F. Asadi2, H. Biranvand3, P. Banaei4, M. Torkashvand5, A. Nazari1, M. Rafieian-Kopaei6,
P. Hashemzadeh7, A.A. Kiani8, V. Ghorbanzadeh1,*
1Cardiovascular Research Center, Shahid Rahimi Hospital, Lorestan University of Medical Sciences, 2Lorestan
University of Medical Sciences, Faculty of Pharmacy, 3Lorestan University of Medical Sciencse, Department of
Physiology, Khorramabad, 4Bu Ali Sina University, Faculty of Sports Sciences, Hamadan, 5University of Tehran,
College of Engineering, Fouman Faculty of Engineering,Tehran, 6Shahrekord University of Medical Sciences,
Basic Health Sciences Institute, Medical Plants Research Center, Shahrekord, 7Lorestan University of Medical
Sciences, Department of Medical Biotechnology, Faculty of Medicine, Khoramabad, 8Lorestan University of
Medical Sciences, Department of Hematology and Blood Transfusion, Khoramabad, Iran
*Correspondence to: Vajihe Ghorbanzadeh Ph.D, Cardiovascular Research Center, Shahid Rahimi Hospital, Lorestan University of
Medical Sciences, Khoramabad, Iran, E-mail: vghorbanzadeh@gmail.com
Abstract
Background. Valepotriate is an active ingredient
of valerian (Valeriana officinalis) with strong antioxidant
activity that is effective for numerous cardiovascular
diseases.
Objective. The aim of this study was to investigate
the effect of an active ingredient of V. officinalis extract on
ischemia-reperfusion-induced cardiac injuries in male rats.
Methods. Thirty-two male rats were subjected to
ischemia for 40 minutes and reperfusion for five days. The
rats were divided into 4 groups of 8 each; group 1 (control)
was given normal saline, and groups 2-4 were gavaged with
0.2, 0.1, 0.05 mg/kg of valepotriate extract, respectively, and
received extract (0.2 mg/kg ip) two weeks before ischemia
induction.
Results. Dichloromethane V. officinalis
(valepotriate) extract exerted a protective effect against
ischemia-reperfusion-induced injuries. So that infarct size
and number of ventricular arrhythmia and ventricular escape
beats decreased compared to the control group. Moreover, ST
segment amplitude, QTC interval, and heart rate decreased
in the injured hearts and serum levels of antioxidant enzymes
glutathione peroxidase, catalase, and superoxide dismutase
increased. Biochemical markers malondialdehyde and
lactate dehydrogenase also decreased on day 5 after the onset
of reperfusion.
Conclusion. V. officinalis extract may have a
protective effect against myocardial ischemia-reperfusion by
producing antioxidant effects.
Keywords: Valepotriate, Antioxidants, Ischemia,
Reperfusion, Cardiac injurey, ST segment, QTC interval.
INTRODUCTION
Cardiovascular diseases (CVDs) including
coronary artery disease (CAD), hypertension,
congenital heart disease and myocardial infarction
(MI) are the leading cause of death internationally (1).
The reasons for CVDs include hypertension, hypoxia,
elevated total cholesterol and low-density lipoprotein
(LDL), decreased serum high-density lipoprotein
(HDL) level, diabetes mellitus, and aging. Oxidative
stress and inflammation are also important risk
factors for CVDs (2). Cardiac ischemia causes tissue
damage and dysfunction by increasing the production
of free radicals, nitric oxide (NO) and decreasing the
activity of antioxidant enzymes such as superoxide
dismutase (SOD), glutathione peroxidase (GPO), and
catalase (CAT) (3). MI is the most common cause of
heart failure. MI is associated with changes such as
cardiomyocyte hypertrophy, myocardial arrhythmia,
left ventricular systolic and diastolic dysfunction,
decreased left ventricular contractile strength, increased
fibrosis and apoptosis, and decreased capillary density
(4). Inducing reperfusion in the ischemic heart is one
of the therapeutic methods that, in addition to assisting
in treatment, can cause a widely varied tissue damage
(5,6). On the other hand, reperfusion potentially might
lead to lethal ventricular arrhythmias (VAs) such as
tachycardia and ventricular fibrillation (VF) (7).
One of the known ways to reduce the side effects
1300

Cardioprotective effect of Valeriana officinalis
179
of reperfusion is preconditioning. Preconditioning
(PC) refers to the preparation of the tissue using a
physical stimulus such as inducing short-term ischemic
durations [ischemic PC (IPC)] or pharmacological
stimulus before induction of ischemia-reperfusion,
so that tissue resistance is increased and the severity
of ischemia-reperfusion-induced tissue injuries are
decreased by stimulating and creating endogenous
defense mechanisms (8).
Today, CVD imposes a huge economic burden
on society and patients and many companies invest
numerous resources on new drugs development for
these diseases. The medicinal plants, besides ischemia-
reperfusion, could be considered as a good candidate
for CVD treatment with low cost and fewer side effects
(9). Studies demonstrates that, antioxidants play an
important role in creating IPC properties. Antioxidants
by scavenging oxygen-derived free radicals have a
stunning effect on cardiac contractility (9).
Among the most popular herbal medicines
used for their antioxidant effects, different species
belonging to the “Valerianaceae” family are in the
spotlight of research. Valeriana officinalis is one of the
members of this family and consists of about 150-200
chemical constituents including flavonoids, triterpenes,
lignans and alkaloids but the roots and rhizomes of
this plant, consist of two main groups of compounds
include sesquiterpenes of volatile oil and valepotriates
(10,11). In ancient medicine, Valerian exhibits
vasodilatory and antiarrhythmic properties. Studies
revealed that these properties effects on mice, rabbits
and cats are associated with valepotriates (12). In this
study we evaluate the impact of preconditioning with
valepotriate extract on ischemia-reperfusion-induced
cardiac injuries in male rats.
MATERIALS AND METHODS
Animal models
In this experimental study, 40 male Wistar rats
(assigned to 5 groups of 8 each) weighing 250-300 g
were used. The rats had ad libitum access to water and
food under standard laboratory conditions [12-hour
light/12-hour dark cycle and 22±2˚C temperature].
Preparation of Valeriana officinalis extract
Following described protocol by Backlund et
al. in 1998 we extracted valepotriates from V. officinalis
rhizomes with some modifications. V. officinalis
rhizomes was purchased from a grocery in the center
of Lorestan province and then shade dried at 20˚C
and powdered. Twenty grams of rhizome powder was
extracted with 300 mL of dichloromethane solvent for
24 hours and then filtered. The liquid under the filter
was evaporated using a rotary apparatus under vacuum
at 30 ˚C, and the resulting dichloromethane extract was
wrapped in foil and stored in the refrigerator at 4˚C for
the next steps. Extraction using dichloromethane causes
the extraction of various derivatives of valepotriate
from plant rhizomes (13).
Experimental design
1. Control group: given normal saline by
gavage (Control)
2. Valepotriate extract at 0.05 mg/kg by gavage
(VT0.05)
3. Valepotriate extract at 0.1 mg/kgby gavage
(VT0.1)
4. Valepotriate extract at 0.2 mg/kg by gavage
(VT0.2)
5. Valepotriate extract at 0.2 mg/kg by
intraperitoneal injection (IP) (VT0.2+ IP).
Animal preparation
The animal was first anesthetized by IP sodium
thiopental (60 mg/kg), its neck and chest were shaved,
and then it was placed on the surgical table. A small
lamp was placed on the surgical table to keep the
animal's body temperature at approximately 37˚C.
The animal was then connected to a small animal
ventilator [Harvard Model 683-USA (60-70 breaths
per minute and current volume of 15 ml/kg)] and saliva
was suctioned. The electrodes were then connected to
record the lead II electrocardiogram of the PowerLab.
Inducing a temporary MI
First, an incision was made between the third
and fourth ribs of the chest to expose the heart. It should
be noted that the incision should be carefully made so as
not to damage the lungs or heart. The pericardium was
then gently ruptured and the 0.6 silk thread was carefully
passed under the left anterior descending (LAD) artery.
The coronary artery closed approximately 2 mm below
the left atrium. Loosening the knot and pulling the suture
induces temporary ischemia. Rats were subjected to
ischemia for 40 minutes and reperfusion for five days.
Electrocardiography changes were monitored with the
PowerLab to ensure the infarction in rats. The layers of
muscle and skin were then sutured. At the completion
of the procedure, the animal was placed under pure
oxygen to regain consciousness. The animal's body
temperature was maintained at 37±1˚C using a heat pad

M. Sedighi et al.
180
during the procedure. After the animal regained full
consciousness, it was transferred to the cage, provided
with water and food, and taken to an animal house.
Determining infarct size
At the end of the study and completion of
reperfusion period, the heart was removed under deep
anesthesia. First, the heart was rinsed with distilled
water, and its atria, vascular roots and additional
appendages were separated, and then it was wrapped
in an aluminum foil and left in the freezer until the
next day. After removing the heart from the freezer, 2
mm sections of the left ventricle (from the base to the
apex) were prepared using graduated molds. Next, they
were stained with 2,3,5-triphenyltetrazolium chloride
at 37˚C for 20 minutes, fixed in formalin 10% and then
the percentage of infarct size was calculated by image
J software (National Institute of Health, Bethesda, MD,
USA).
Measurement of cardiac enzymes and
antioxidants
At the end of reperfusion, blood samples
were collected from the carotid artery and centrifuged
at 5000 rpm for 15 minutes. The serum was frozen
and stored at -70˚C until subsequent measurements.
Finally, as the markers of myocyte necrosis, the serum
levels of lactate dehydrogenase (LDH) were measured
by LDH assay kit (Pars Azmoon Co., Iran) and
troponin I (CTnI) measured by Zellbio kit (Germany)
via autoanalyzer apparatus (Roche Hitachi Modular DP
Systems, Mannheim, Germany).
Plasma MDA level, as an indicator of oxidative
stress, was measured based on thiobarbituric acid
reaction by spectrophotometry method. Measurements
of antioxidant enzymes include SOD, GPx, and CAT
were performed according to the kit's instructions and a
spectrophotometer apparatus (Shimadzu, Tokyo, Japan)
(All kits provided from Pars Azmoon Co., Iran) (14).
RESULTS
Effect of V. officinalis extract on serum
antioxidant enzymes activity
The level of SOD activity in serum is shown
in Figure 1a. In comparison to the control group, SOD
activity significantly increased after administration of
V. officinalis extract in VT0.1, VT0.02 and VT0.2 IP
groups in dose dependent manner (P<0.01). In VT0.2
group SOD activity increased 2.1-fold rather than
control group but in inter peritoneal injected group
SOD showed 0.4-fold lower activity.
Figure 2b showed activity of GPX. In
comparison with control group, GPX activity increase
significantly in VT0.2 group (1.9-fold).
P<0.05 is considered as signicant
A
B
C
Figure 1. Eect of Valeriana ocinalis extract on oxidave stress
markers. Results are expressed as mean±SEM. (a) glutathione
peroxidase (GPx), (b) superoxide dismutase (SOD), (c) catalase
(CAT), (d) malondialdehyde (MDA). VT0.05, V. ocinalis extract
at 0.05mg/kg; VT0.1, V. ocinalis extract at 0.1 mg/kg; VT0.2,
V. ocinalis extract at 0.2 mg/kg; VT0.2 IP, V. ocinalis extract
at 0.2 mg/kg by peritoneal injecon. *P<0.05 and ***P<0.001,
signicant dierence compared to control group; ##P<0.01 and
###P<0.001, signicant dierence compared to VT0.05 group;
$$P<0.05, signicant dierence compared to the VT0.1 group;
@@P<0.01 and @@@P<0.001, signicant dierence compared to
the VT0.2 IP group.

Cardioprotective effect of Valeriana officinalis
181
As you seen in Figure 2c, CAT activity
increases 1.6-fold in 0.2VT group rather than control.
V. officinalis extract alter biochemical factors
The serum LDH, MDA and cTnI levels in
rats in different groups were shown in Figure 2. Five
days after reperfusion, rather the control group cTnI
decreased in all 4 groups but this reduction is not
statistically significant (P>0.05).
As seen in Figure 2b, LDH in animals that
received 0.2 mg/kg V. officinalis extract significantly
decreased more than 2-fold. MDA in Figure 2c exhibit
same manner and decrease 2.2 and 2.7-fold in 0.2 VTIP
and VT0.1 respectively.
Infarct size of myocardial tissue
As illustrated in Figure 3, treatment with V.
officinalis extract caused a significant reduction in the
percentage of infarcted tissue at VT0.1, VT0.2 and
VT0.2 IP compared to the control group (P<0.001,
P<0.01 and P<0.01, respectively) (Fig. 3).
Determination and evaluation of ventricular
arrhythmias (VAs)
In this study, VAs including ventricular
escape beats (VEBs), ventricular tachycardia (VT)
and ventricular fibrillation (VF) were evaluated based
on the Lambeth model. VEB refers to premature and
flattened QRS complexes that are called bigeminy
when they occur one in between with the normal QRS
complex and are called trigeminy when one VEB occurs
for both natural QRS complexes. In other combination
types such as couplets (two VEBs in a row) and triplets
(three VEBs in a row), VEBs were counted separately.
VT refers to the generation of more than three
VEBs in a row. VFs refer to indeterminate and low
voltage QRS complexes that, if they elapse for less than
2 minutes, will be transient, and otherwise, they will be
A
B
C
Figure 2. Eect of Valeriana ocinalis extract on biochemical
markers. Results are expressed as Mean±SEM. (A) troponin I (cTnI),
(B) lactate dehydrogenase (LDH), (C) malondialdehyde (MDA).
VT0.05, V. ocinalis extract at 0.05 mg/kg; VT0.1, V. ocinalis
extract at 0.1 mg/kg; VT0.2, V. ocinalis extract at 0.2 mg/kg by
gavage; VT0.2 IP, V. ocinalis extract at 0.2 mg/kg by peritoneal
injecon; **P<0.01, signicant dierence compared to the control
group.
Figure 3. Eect of valepotriate extract on infarct size percentage.
The results are expressed as mean±.SEM. VT0.05, valepotriate
extract at 0.05 mg/kg; VT0.1, valepotriate extract at 0.1 mg/kg;
VT0.2, valepotriate extract at 0.2 mg/kg; **P<0.01 and ***P<0.001,
signicant dierence compared to the control group.

M. Sedighi et al.
182
stable. In this study, the incidence of VF, duration and
number of VTs, as well as the severity of Vas, were
investigated at 30-minute ischemia intervals.
Regarding the number of VEBs, treatment
with V. officinalis extract at 0.1 mg/kg (VT0.1) and
0.2 mg/kg (VT0.2) significantly reduced the number
of arrhythmias at the 30-minute ischemia interval
compared to the control group (P<0.01 for both) (Fig.
4a). Furthermore, the number of VTs observed at the
30-minute ischemia interval significantly decreased
at 0.2 mg/kg (VT0.2) compared to the control group
and at 0.05 mg/kg (VT0.05) and 0.1 mg/kg (VT0.1)
(P<0.05 for all, Fig. 4b).
Electrocardiography parameters
HR
HR at baseline interval was significantly different
between all groups. Treatment with V. officinalis extract
significantly reduced the number of HR at 0.1 mg/kg
(VT0.1) at the 30-minute ischemia interval compared
to the control group (P<0.05). Furthermore, the number
of HR at 60-minute reperfusion interval at 0.1 mg/kg
(VT0.1) and 0.2 mg/kg (VT0.2) significantly decreased
compared to the control group (P<0.05). Comparison
of different intervals of the control group showed
that the 30-minute ischemia interval and 60-minute
reperfusion interval significantly increased compared
to the baseline (P<0.01 and P<0.001, respectively).
Besides that, the 60-minute reperfusion interval in
the control group significantly increased compared
to the 30-minute reperfusion interval (P<0.05). After
intraperitoneal injection of 0.2 mg/kg extract (VT0.2
IP), the 60-minute reperfusion interval significantly
increased compared to the baseline (P<0.05, Table 1).
QTc length (ms)
The duration of QTc in different groups was
not significantly different at baseline. Pretreatment
with V. officinalis extract at the 30-minute ischemia
interval significantly decreased during QTc at 0.05 mg/
kg (VT0.05) and 0.1 mg/kg (VT0.1) compared to the
control group (P<0.05 and P<0.01, respectively). QTc
duration at the 60-minute reperfusion interval at 0.05
mg/kg (VT0.05) and 0.01 mg/kg (VT0.1) significantly
Figure 1. A: Before treatment; B: Aer treatment.
End of reperfusion 60’End of ischemia 30’BaselineGroups
311.08
±13.59###$
270.40±7.98##
211.15±10.56
Control
253.84
±8.83
239.79
±16.86213.53±17.85
VT 0.05
249.77
±16.17*
205.11
±15.55*201.4±13.25
VT 0.1
243.19
±16.60*220.54±13.31215.23±16.05
VT 0.2
285.44±#18.65249.8±14.29223.05±14.44
VT 0.2 IP
Table 1. Eect of valepotriate extract on heart rate in the last ve days of reperfusion
Results are expressed as Mean±SD. VT0.05, valepotriate extract at 0.05 mg/kg; VT 0.1, valepotriate extract at 0.1 mg/kg; VT0.2, valepotriate extract
at 0.2 mg/kg; VT0.2 IP, intraperitoneal injecon of valepotriate extract at 0.2 mg/kg. *P<0.05, signicant dierence compared to the control group in
the corresponding interval; #P<0.05, signicant dierence compared to baseline; ##P<0.01, signicant dierence compared to baseline; ###P<0.001,
signicant dierence compared to baseline; $P<0.05, signicant dierence compared to 30-minute reperfusion interval.
Figure 4. Eect of valepotriate extract on the number of arrhythmias recorded on electrocardiography. Results are expressed as
Mean±SEM. (a) Number of ventricular escape beats (VEBs); (b) Number of ventricular tachycardia (VT); VT0.01, valepotriate extract
at 0.1 mg/kg; VT0.2, valepotriate extract at 0.2 mg/kg; VT0.2 IP, intraperitoneal injecon of valepotriate extract at 0.2 mg/kg; *P<0.05
and **P<0.01, signicant dierence compared to control group; #P<0.05, signicant dierence compared to VT0.05 group; $P<0.05,
signicant dierence compared to VT0.1 group.

Cardioprotective effect of Valeriana officinalis
183
decreased compared to the control group (P<0.05 for
both). In comparison to the control group at different
intervals, the 30-minute ischemia interval and the
30-minute reperfusion interval significantly increased
compared to the baseline interval (P<0.001 and P<0.01,
respectively) (Table 2).
R wave amplitude (μv)
R wave amplitude was not significantly
different at baseline in different groups and at the
60-minute reperfusion interval in the control group and
the VT0.2 IP group (0.2 mg/kg), but treatment with
V. officinalis extract at 0.1 mg/kg (VT0.1) reduced R
wave amplitude compared to 0.2 mg/kg (VT0.2) at the
30-minute ischemia interval (P<0.01). The comparison
of the intervals showed that the R wave amplitude at
0.05 mg/kg (VT0.05), 0.1 mg/kg (VT0.1) and 0.2 mg/
kg (VT0.2) significantly decreased at the 60-minute
reperfusion interval compared to the baseline interval
(P<0.05, P<0.01 and P<0.05, respectively) (Table 3).
T wave amplitude (μv)
T wave amplitude in different groups was not
significantly different at baseline, 30-minute ischemia
interval and 60-minute reperfusion interval, but
significantly decreased at the 60-minute reperfusion
interval after intraperitoneal injection of 0.2 mg/kg
(VT0.2 IP) compared to the baseline interval and at the
corresponding dose (P<0.05, Table 4).
ST-segment amplitude (μv)
ST-segment amplitude in different groups was
not significantly different at baseline. Pretreatment with
V. officinalis extract reduced ST segment amplitude at
the 30-minute ischemia interval at 0.1 mg/kg (VT0.1)
and after intraperitoneal injection of 0.2 mg/kg (VT0.2
IP) compared to the control group. In addition, ST-
segment amplitude significantly decreased at the
60-minute reperfusion interval at 0.05 mg/kg (VT0.05),
0.1 mg/kg (VT0.1) and after intraperitoneal injection
of 0.2 mg/kg (VT0.2 IP) compared to the control
group (P<0.05, P<0.01 and P<0.05, respectively).
Comparison between different intervals in the control
group showed that the 30-minute ischemia interval
significantly decreased compared to the baseline
interval (P<0.05). Moreover, the extract at 0.2 mg/
kg by gavage (VT0.2) significantly increased at the
End of reperfusion 60’End of ischemia 30’BaselineGroups
0.338
±0.093
0.477
±0.066
0.519
±0.05
Control
0.337
±0.079#
0.511
±0.034
0.509
±0.09
VT 0.05
0.264
±0.053##
0.335
±0.026**
0.516
±0.041
VT 0.1
0.450
±0.084#
0.581
±0.051
0.634
±0.030
VT 0.2
0.482
±0.041
0.513
±0.018
0.519
±0.041
VT 0.2 IP
The results are expressed as mean±SD. VT 0.05, valepotriate extract at 0.05 mg/kg; VT0.1, valepotriate extract at 0.1 mg/kg; VT0.2, valepotriate extract
at 0.2 mg/kg; VT0.2 IP, intraperitoneal injecon of 0.2 mg /kg valepotriate extract. **P<0.01, signicant dierence compared to the control group at the
corresponding interval; #P<0.05 and ##P<0.01, signicant dierence compared to the baseline interval.
Table 3. R wave amplitude on the last ve days of reperfusion
Groups Baseline End of ischemia 30’ End of reperfusion 60’
Control 0.0907±0.0226 0.1748±0.0709 0.0734±0.0294
VT 0.05 0.1103±0.0175 0.1716±0.0236 0.0704±0.0387
VT 0.1 0.1407±0.0224 0.1484±0.0266 0.0531±0.0309
VT 0.2 0.1529±0.0094 0.1713±0.0220 0.1153±0.0191
VT 0.2 IP 0.1364±0.0084 0.1184±0.0747 0.0469±0.0160#
The results are expressed as Mean±SD. VT0.05, valepotriate extract at 0.05 mg/kg; VT0.1, valepotriate extract at 0.1 mg /kg; VT 0.2, valepotriate extract
at 0.2 mg/kg, VT0.2 IP; intraperitoneal injecon of 0.2 mg/kg valepotriate extract. **P<0.01, signicant dierence compared to the control group at the
corresponding interval. #P<0.05, signicant dierence compared to the control group and at the corresponding dose.
Table 4. T wave amplitude on the last ve days of reperfusion
End of reperfusion 60’End of ischemia 30’BaselineGroups
0.184
±0.019##
0.197
±0.007###
0.113
±0.003
Control
0.119
±0.010*
0.152
±0.011*
0.123
±0.013
VT 0.05
0.124
±0.012*
0.138
±0.015**
0.121
±0.08
VT 0.1
0.138
±0.009
0.158
±0.009
0.130
±0.010
VT 0.2
0.129
±0.011
0.159
±0.009
0.125
±0.013
VT 0.2 IP
Results are expressed as Mean±SD. VT0.05, valepotriate extract at 0.05 mg/kg; VT0.1, valepotriate extract at 0.1 mg/kg; VT0.2, valepotriate extract at 0.2
mg/kg; VT0.2 IP, intraperitoneal injecon of valepotriate extract at 0.2 mg/kg. *P<0.05 and **P<0.01, signicant dierence compared to control group at
the corresponding interval; ##P<0.01 and ###P<0.001, signicant dierence compared to the baseline interval.
Table 2. QTc alteraons in the last ve days of reperfusion

M. Sedighi et al.
184
30-minute ischemia interval compared to the baseline
and 60-minute reperfusion intervals (P<0.05 for both,
Table 5).
Electrocardiography parameters
Left ventricular end-diastolic volume (LVS) at
0.2 mg/kg (VT0.2) significantly increased compared to
the control group (P<0.05), but for other parameters,
no significant difference was observed between the
treatment groups and the control groups (Table 6).
DISCUSSION
This study showed that dichloromethane V.
officinalis (valproate) extract has a protective effect
against ischemia-reperfusion, which was confirmed by
the reduction of infarct size and the number of VTs and
VEBs in comparison with the control group. The extract
also caused a decrease in ST segment amplitude, QTc
interval and the number of HR in the affected heart,
an increase in the serum levels of antioxidant enzymes
GPx, CAT and SOD, and a decrease in biochemical
markers include cTnI, MDA and LDH on day 5 after
the onset of perfusion.
For over two decades, the role of oxygen species
in heart disease has been studied and findings revealed
that there is a balance between the forms of oxygen
radicals and antioxidants amount in normal conditions.
In some pathophysiological conditions and subsequent
reperfusion, disconnection and loss of balance may
occur, but the products of free radicals are reduced in
the presence of antioxidants (15). ROS play a pivotal
role in creating ischemia-reperfusion-induced injuries
(16). ROS cause a number of disorders including
cardiomyocyte hypertrophy, apoptosis, fibrosis and
decreased capillary density, arrhythmia, systolic
and diastolic dysfunction, and decreased ventricular
contractile strength in the myocardium in vitro and
in vivo (4). According to the previous studies, ROS
have a substantially destructive effect on myocardial
contractility, which can be prevented during ischemia-
reperfusion by antioxidants pretreatment (17,18).
Beneficial effects of long-term use of antioxidants in
preventing heart failure after reperfusion by inhibiting
apoptosis have already been reported (19).
Numerous studies have been performed in
different laboratories that show the positive effect of
CAT and SOD in ischemia-reperfusion models (20,21).
The antioxidant effects of valerian have been confirmed
in previous studies (22,23). It has been observed that
valerian extract can activate and induce the antioxidant
enzymes SOD and GPx (24). Wang et al. in 2017
has shown that valepotriates in dichloromethane V.
officinalis extract reduce the amount of ROS and have
strong antioxidant effects (25). Also, it has been shown
that valepotriates can exert antioxidant properties
via the GABA signaling pathway (26,27). Based on
research evidence, valepotriates reduce enzymes and
biomarkers such as xanthine oxidase, LDH, MDA and
tumor necrosis factor (24). It seems that the protective
effect of V. officinalis extract in previous studies might
End of reperfusion 60’End of ischemia 30’BaselineGroups
0.148
±0.013
0.182
±0.004##
0.074
±0.032
Control
0.037
±0.036*
0.099
±0.030
0.067
±0.020
VT 0.05
0.001
±0.024**
0.051
±0.015*
0.066
±0.021
VT 0.1
0.063
±0.01
0.136
±0.027#$
0.061
±0.009
VT 0.2
0.037
±0.033*
0.067
±0.048*
0.021
±0.02
VT 0.2 IP
Table 5. ST-segment amplitude on the last ve days of reperfusion
Results are expressed as mean±SD. VT0.05, valepotriate extract at 0.05 mg/kg; VT0.1, valepotriate extract at 0.1 mg/kg; VT0.2, valepotriate extract at 0.2
mg/kg, VT0.2 IP; intraperitoneal injecon of 0.2 mg /kg valepotriate extract. *P<0.05 and **P<0.01, signicant dierence compared to the control group
at the corresponding interval; #P<0.05 and ##P<0.01, signicant dierence compared to baseline; $P<0.05, signicant dierence compared to 60- minute
reperfusion interval.
VT 0.2 IPVT 0.2VT 0.1VT 0.05ControlShamGroup
1.645±0.1422.204±0.032* 2.193±0.1052.123±0.2441.60±0.0592.10±0.287LVS(mm)
4.040±0.0634.100±0.2043.850±0.0574.162±0.3493.682±0.1824.150±0.285LVD(mm)
0.358±0.2350.127±0.0050.162±0.0100.127±0.0130.373±0.1310.119±0.006SH.TIME(ms)
1.160±0.1031.299±0.0691.269±0.1441.483±0.1441.029±0.1161.170±0.084SEPD(mm)
82.795±3.09669.725±3.36066.570±5.97575.809±4.66279.813±2.45771.895±7.079FS
59.183±3.693*45.440±2.90742.915±5.30151.823±4.95155.625±2.87448.625±6.539EF
Table 6. Electrocardiography parameters alteraons
Results are expressed as mean± SD. LVS, le ventricular end-systolic volume; LVD, le ventricular end-systolic volume; SH.me, le ventricule lling up and
emptying me; SEPD, the thickness of the wall between the two ventricles (calculated by the formula ); FS, fraconal shortening;
EF, ejecon fracon (calculated by the formula *P<0.05, signicant dierence compared to the control group.

Cardioprotective effect of Valeriana officinalis
185
be related to antioxidant content of this plant. It has
been observed that valepotriates in V. officinalis extract
have antiarrhythmic properties (28-30).
Previous studies have also shown the
vasodilation and vasorelaxation effects of valepotriates
(24,28,31,32). In our study, the number of VEBs at
0.1 mg/kg and 0.2 mg/kg extract and the number of
VTs at 0.2 mg/kg extract decreased compared to the
control group, which confirms the antiarrhythmic
effects of valepotriates at the 30-minutes ischemia
interval. Valepotriates have positive inotropic and
negative chronotropic effects (12). Here, HR at the
30-minute ischemia interval significantly increased
compared to baseline, but in comparison with the
30-minute ischemia interval, a significant decrease was
observed at 0.1 mg/kg extract compared to the control
group, which could confirm the negative chronotropic
properties of valepotriate.
Potassium channels play an important role in
stimulating the heart and usually serve as a target in
determining the effect of antiarrhythmic compounds,
and prevent QTc from rising, which results in
arrhythmia and death (33,34). Therefore, development
of antiarrhythmic therapeutics for reducing QTc has
been one of the main goals of many studies. In the
present study, a decrease in QTc interval was observed
at 0.1 mg/kg and 0.2 mg/kg extract at the 30-minute
ischemia and 30-minute reperfusion intervals.
R wave amplitude is one of the
electrocardiography parameters of myocardial
contractility (35). ST segment and T wave amplitudes
are other important markers of ischemia. T-wave is one
of the electrocardiography parameters and indicates
ventricular repolarization. Decreased T-wave amplitude
helps to diagnose heart disease (36). In the current
study, ST segment amplitude at 0.1 mg/kg extract and
after intraperitoneal injection of 0.2 mg/kg extract
significantly decreased compared to the control group
at the 30-minute ischemia interval, but the amplitude
of R wave and T-wave did not significantly increase
in different groups. Therefore, more extensive studies
should be done to determine the effects of valepotriate
on the electrical activity of the heart.
In our study, LVS significantly increased
compared to the control group, but other
electrocardiography parameters did not change
significantly. The level of ROS increases after the
occurrence of ischemia-reperfusion, while ROS
decreases if flavonoids are pretreated before ischemia-
reperfusion occurs (37). Furthermore, the level of
SOD increases after pretreatment compared to before
pretreatment and the level of MDA decreases.
In conclusion, given our results and previous
studies, it can be concluded that dichloromethane
extract of V. officinalis rhizome, which mainly
contains valepotriates, exerted favorable impacts
on antioxidant enzymes (MDA, GPx, SOD and
CAT), biochemical factors, infarct size and
electrocardiography parameters.
Taken together, valepotriate extract mitigated
injuries due to before induction of ischemia-
reperfusion. Our study, as with research, showed the
strong antioxidant activity of the extract and the role of
alkaloids such as valepotriate. Therefore, valerian with
strong antioxidant properties may play an important
role in preventing cardiac injuries such as arrhythmia
and infarct size, and also in activating antioxidant
enzymes in heart tissue.
Conflict of interest
The authors declare that they have no conflict of
interest.
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