Abstract Oxidative stress plays a major role in biochemical and pathological changes associated with myocardial ischemic-reperfusion injury (IRI).The need to specify agents with a potential for preventing such damage with therapeutic importance. In the present study, chronic oral. International Journal of Medicinal Plants. Photon 107 (2014) 572-579 https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-medicinal-plants Original Research Article. ISJN: 6672-4384: Impact Index: 3.12

1. International Journal of Medicinal Plants. Photon 107 (2014) 572-579
https://sites.google.com/site/photonfoundationorganization/home/international-journal-of-medicinal-plants
Original Research Article. ISJN: 6672-4384: Impact Index: 3.12
International Journal of Medicinal Plants Ph ton
Therapeutic response of Mimosa pudica in oxidative stress induced by
ischemic reperfusion injury on rat heart
Sudha Kumaria, Muralidhar S. Talkadb*, Chethan Chowdharyb, Kavya S.b, Parvathi C.b, Aamir Javedc,
Umesh H.R.d, Sivakami Sundari P.e
a CMJ University, Shillong, India
b P.G. Department of Biotechnology, R&D Centre, Dayananda Sagar College of Biological Sciences,
Kumaraswamy Layout, Bangalore-560078, India
c Base Fertility Medical Science Pvt. Ltd. Bangalore- 560079 India
d P.G. Department of Biochemistry
e Department of Pharmacognosy, Dayannanda Sagar College of Pharmacy, Kumaraswamy Layout, Bangalore-
560078, India
Article history:
Received: 04 February, 2014
Accepted: 09 February, 2014
Available online: 11 August, 2014
Keywords:
Mimosa pudica, oxidative stress, myocardial ischemic-reperfusion
injury (IRI), TBARS, SOD, catalase
Abbreviations:
IRI: myocardial ischemic-reperfusion injury, IHD: Ischemic
heart disease, TBARS: Thiobarbituric acid reactive substance,
SOD: Superoxide Dismutase, GSH: Reduced glutathione, GPx:
Glutathione peroxidise
Corresponding Author:
Talkad M.S.*
Professor
Email: talkad.murali@rediffmail.com
Kumari S.
Research Scholar
Javed A.
Jr. Embryologist
Sundari S.P.
Assistant Professor
Abstract
Oxidative stress plays a major role in biochemical
and pathological changes associated with
myocardial ischemic-reperfusion injury (IRI).The
need to specify agents with a potential for
preventing such damage with therapeutic
importance. In the present study, chronic oral
administration of Mimosa pudica plant extract on
oxidative stress induced by ischemic-reperfusion
injury in isolated rat heart was investigated. Mimosa
pudica plant extract (125, 250, & 500 mg/kg once
daily for 30 days) was administered orally in Wistar
albino rats. Thereafter, hearts were isolated and
subjected to IRI. Depletion of myocardial
endogenous antioxidants and rise in TBARS were
significantly less in the Mimosa pudica plant extract
treated rat hearts. Oxidative stress induced cellular
damage as indicated by patchy necrosis,
inflammatory cells and marked interstitial edema in
the muscle fibres were noted in Control –IR group
but in M-500 group the absence of myonecrosis
was observed. Since Mimosa pudica plant extract
possess strong antioxidant activities in one of our
earlier studies followed by this study strongly
suggests that protective therapeutic response in the
same plant extract at the dose of 500 mg/kg/day for
30 days, hence the Mimosa pudica plant extract
prevents oxidative stress and associated structural
changes induced by myocardial ischemic-reperfusion
injury.
Citation:
Kumari S., Talkad M.S., Chowdhary C., Kavya S., Parvathi C.,
Javed A., Umesh H.R., Sundari S.P., 2014. Therapeutic
response of Mimosa pudica in oxidative stress induced by
ischemic reperfusion injury on rat heart. International Journal of
Medicinal Plants. Photon 107, 572-579.
All Rights Reserved with Photon.
Photon Ignitor: ISJN66724384D656811082014
1. Introduction
Oxidative stress is a well established
etiopathogenic factor of ischemic heart disease
(IHD) and its consequences. Reperfusion of the
ischemic myocardium is only logical approach for
the successful management of patients with acute
obstructions of the ischemic myocardial tissue
undergoing reperfusion injury is a true pathological
phenomenon and a distinct entity from the
preceding ischemic injury.
Generation of reactive oxygen species immediately
upon reperfusion has been documented in
Ph ton 572

2. experimental conditions, as well as in patients with
acute myocardial infarction undergoing
thrombolysis, coronary angioplasty or open heart
surgery. Upon reperfusion, molecular oxygen
undergoes sequential reduction to form reactive
oxygen species, including superoxide anion and
hydroxyl radical, in addition to hydrogen peroxide.
The interaction of oxygen derived free radicals
with cell membrane lipids and essential proteins
contribute to myocardial cell damage, leading to
depressed cardiac function and irreversible tissue
injury with concomitant depletion of certain key
endogenous antioxidant compounds, e.g.,
superoxide dismutase (SOD), catalase, reduced
glutathione (GSH) and Glutathione peroxidise
(GPx)
Although antioxidant administration during
reperfusion injury has been shown to reduce the
severity of the ischemic reperfusion injury, yet
some properties of antioxidants such as
cytotoxicity, pro-oxidant activity or high molecular
weight in case of SOD, limited their therapeutic
application.
2. Objective of Research
Use of plant extracts, food supplement and even
drugs which augment major cellular endogenous
antioxidants following chronic administration have
been identified as a promising therapeutic approach
to combat oxidative stress associated with ischemic
heart disease. The property of augmenting cellular
endogenous antioxidants has been defined as a
major constituent of myocardial adaptation against
oxidative stress.
Some experimental studies have claimed that
Mimosa pudica plant extract against oxidative
stress exhibited beneficial Antioxidant properties
(Zuhaib et al., 2011) and it could show significant
beneficial effects in ischemic heart disease (IHD).
Since Garlic reduces a multitude of risk factors
which play a decisive role in the genesis and
progression of IHD, e.g., hypolipidemic effect,
lowering of arterial blood pressure and inhibition of
platelet aggregation. Recently, it has been shown
that chronic administration of garlic augmented
SOD and catalase in rat heart. In view of this
observation, the present study was designed to
investigate cytoprotection of Mimosa pudica plant
extract against oxidative stress arising out of
ischemic-reperfusion injury in isolated rat heart.
2.1 Brief summary of work plan
Selection on whole plant Mimosa pudica –
subjected for methanolic extract in Soxlet
apparatus - The yield of dried extract was
approximately 40% and a homogenate was made in
CMC - Pre treatment with extract at 3 different
dose limits, 0.05, 0.1 and 0.2 gm/ml, corresponding
to 125 mg, 250 mg and 500 mg/kg body weight of
animal – for 30 days, Since Group C: Normal
saline-fed rat hearts subjected to 26-min.perfusion.
Group C-IR: Normal saline-fed rat hearts subjected
to IR injury. Group M-125: 125 mg/kg/day for 30
days of Mimosa pudica plant extract administered
rat hearts subjected to IR injury. Group M-250: 250
mg/kg/day for 30 days of Mimosa pudica plant
extract administered rat hearts subjected to IR
injury. Group M-500: 500 mg/kg/day for 30 days
of Mimosa pudica plant extract administered rat
hearts subjected to IR injury.
Experimental rationale - Induction of ischemic-reperfusion
injury in isolated rat heart, followed by
myocardial tissue were subjected for biochemical
estimations, Myocardial tissues examined under a
light microscope for Histopathological profiles, the
need for the study is to establish protective effect of
Mimosa pudica, when it s pre-treated at a regular
dose regimes for 30 days.
2.2 Plant extract
Fresh mature whole plant Mimosa pudica was
collected from natural source in rural parts of South
Karnataka regions of India. Routine
pharmacognostic investigations were carried out to
confirm authenticity of this material. The
identification was carried out in our P.G Dept of
Biotechnology. The whole plant were dried and cut
into small pieces and crushed to a coarse powder
using blender. Coarse powder was subjected to
extraction in Sox let apparatus (in ratio of 1:5 to the
quantity of raw material) for 6 h under reflux
condenser at 45o C using thermostat.
This extract was cooled to room temperature and
evaporated using condenser then filtered through
filter cloth (#100 mesh size), to get methanolic
extract of Mimosa pudica plant . The resulting
extracts were stored in well-packed container at
room temperature for future use. The yield of dried
extract was approximately 40% and a homogenate
was made in CMC. Three different concentrations
of the extract were prepared, 0.05, 0.1 and 0.2
gm/ml, corresponding to 125 mg, 250 mg and 500
mg/kg body weight of animal
2.3 Animals
The study was approved by the Institute Animal
Care Ethics Committee obtained from the Central
Animal facility, Sri Raghavendra Enterprises.
Bangalore. India (Reg. no. 860/6/11/ CPCEA),
were used for the study. Laboratory bred Wistar
albino rats of both sexes weighing between 150 and
200 gm were housed in polypropylene cages at a
population density of six per cage, under controlled
environmental conditions of temperature (27 + 300
Ph ton 573

3. C) with normal pellet diet and water was provided
ad libitum
2.4 Chemicals
All chemicals were of analytical grade and
chemicals required for sensitive biochemical assays
were obtained from Merck Chemicals.
2.5 Experimental protocol
Rats were randomly divided into four groups, each
group having 6 rats. Methanolic extracts of Mimosa
pudica plant extract was fed by oral gavage
everyday at a fixed time (10.00 a.m) for 30 days in
three different doses. Control rats were fed normal
saline daily for 30 days. Changes in body weight,
food and water intake patterns of rats in all groups
were noted throughout the experimental period.
After 48 hours of the last dose, rats were
heparinised (375 units/200 gms i.p), anaesthetized
after 30 min. with sodium pentobarbitone (60
mg/kg i.p) and subjected to the following protocol.
2.6 Production of ischemic-reperfusion injury
in isolated rat heart
Hearts were rapidly excised and washed in ice-cold
saline, and then perfused with the non-recirculating
Langendorff's technique (Hufesco, Hungary)
[(Bolli, 1998; Sanjay et al., 2002) in constant
pressure mode with modified Kreb's Henseleits
buffer containing (in mM): glucose 11.1; NaCl
118.5; NaHCO3 25; KCl 2.8; KH2PO4 1.2;
CaCl2 1.2; MgSO4 0.6, with a pH of 7.4. The buffer
solution equilibrated with 95% O2 + 5% CO2 was
delivered to the aortic cannula at 37°C under 60
mm Hg pressure. Following 5 min-equilibration
period, hearts were subjected to 9 min of zero-flow
[ischemia] and 12 min of re-flow [reperfusion] (IR
injury).
2.7 Group Description
Group C: Normal saline-fed rat hearts subjected to
26-min.perfusion.
Group C-IR: Normal saline-fed rat hearts
subjected to IR injury.
Group M-125: 125 mg/kg/day for 30 days of
Mimosa pudica plant extract administered rat
hearts subjected to IR injury.
Group M-250: 250 mg/kg/day for 30 days of
Mimosa pudica plant extract administered rat
hearts subjected to IR injury.
Group M-500: 500 mg/kg/day for 30 days of
Mimosa pudica plant extract administered rat
hearts subjected to IR injury.
At the end of each experiment, myocardial tissue
was stored in liquid nitrogen for biochemical
estimations, 10% buffered formalin for light
microscopic studies
2.8 Biochemical parameters
Thiobarbituric acid reactive substances (TBARS)
was measured as a marker of lipid peroxidation and
endogenous antioxidants, e.g., superoxide
dismutase (SOD), catalase, reduced glutathione
(GSH) & glutathione peroxidase (GPx) were
estimated.
Light microscopic study: Myocardial tissue was
fixed in 10% formalin, routinely processed and
embedded in paraffin. Paraffin sections (3 μm)
were cut on glass slides and stained with
haematoxylin and eosin (H&E) and examined
under a light microscope for Histopathology
3. Results
3.1 Biochemical parameters
There was no mortality, change in body weight as
well as food and water intake pattern of rats in any
group. The effects of various doses of Mimosa
pudica plant extract on lipid peroxidation and
endogenous antioxidants of ischemic-reperfused
hearts are shown in: Table: 1
3.2 Myocardial TBARS
There was significant increase 248.50 ± 28.22*© (p
< 0.05) in myocardial TBARS in the group C-IR
when compared to the control (C) 200.12 ± 10.02.
Significant decrease in the level of myocardial
TBARS was observed in groups, M-125 (p < 0.02),
M-250 (p < 0.001) and M-500 in comparison to the
C-IR group.
3.3 Myocardial SOD
In C-IR group, there was a significant reduction:
4.62 ± 0.87 (p < 0.05) in myocardial SOD activity
when compared to control (C) group: 5.98 ± 0.64.
Significant increase: 18.24 ± 2.76 (p < 0.001) in
myocardial SOD activities was observed in groups
M-125 and M-250 but not in group M-500 when
compared to C-IR.
3.4 Myocardial Catalase
In the C-IR group there was a significant decrease:
15.38 ± 1.92 (p < 0.001) in myocardial catalase
activity compared to control (C). In the groups M-
125 and M-500, there was no significant increase in
the level of myocardial catalase activities compared
to C-IR. However, significant increase: 46.22 ±
5.34 (p < 0.001) in myocardial catalase activity
was seen only in group M-250.
3.5 Myocardial GSH
Significant decrease: 284.45 ± 25.28 (p < 0.001) in
myocardial GSH level was observed in the group
C-IR in comparison to the control (C): 462.16±
38.64. In the groups M-125, M-250 and M-500,
there was a significant increase: 362.86 ± 17.35#
Ph ton 574

4. and 388.22 ± 12.84# (p < 0.02) of myocardial GSH
levels when compared to C-IR.
3.6 Myocardial GPx
Significant reduction: 264.20 ± 25.14 (p < 0.05) in
GPx activity was observed in C-IR group. No
change of GPx activity was observed in any of the
Mimosa pudica plant extract administrated groups
when compared to C-IR group.
3.7 Light microscopic changes
In Normal Control group - Near Normal cardiac
cell architecture with compact Musculature is
evident. In C-IR group, there was patchy necrosis
of muscle fibers with infiltration of acute and
chronic inflammatory cells are evident. In M-125
group: The degree of myocardial damage with mild
necrosis when compared to C-IR group. In M-250
group, the interstitial edema as well as
inflammation is also moderate to mild. In M-500
group there was focal interstitial edema but no
necrosis was observed.
Table 1: Effect of chronic administration of Mimosa pudica plant extract on TBA-RS, GSH, Catalase, SOD and GPx in rat
heart after ischemic-reperfusion injury
Group TBA-RS
(in mole/gm wet
wt.)
GSH (μg/gm wet
wt.)
CATALASE
(U/mg protein)
SOD
(U/mg protein)
GPx (mU/mg
protein)
C 200.12
± 10.02
462.16
± 38.64
48.52 ± 2.88 5.98 ± 0.64 344.18 ± 38.04
C-IR 248.50
± 28.22*©
284.45
± 25.28©©
15.38 ± 1.92©© 4.62 ± 0.87*© 264.20 ± 25.14*©
G-125 158.24
± 24.02#
362.86
± 17.35#
20.05 ± 5.47 15.66 ± 3.40© 274.82 ± 34.42
G-250 128.68
± 10.34©
388.22
± 12.84#
46.22 ± 5.34© 18.24 ± 2.76© 312.16 ± 48.02
G-500 186.67
± 24.62*
364.20
± 15.49#
11.68 ± 1.84 4.14 ± 0.66 282.28 ± 26.44
All values are expressed as Mean ± SE (n = 6). p values: *© < 0.05, ©©< 0.001 vs Control *< 0.05, # < 0.02, © < 0.001 vs C-IR
(One way ANOVA)
4. Discussion
In the present study, ischemic reperfusion injury
(IRI) was associated with increased oxidative
stress, as evidenced by increase in myocardial
TBARS and depletion of myocardial endogenous
antioxidants such as SOD, catalase, GSH and GPx.
Similar observations were made earlier by different
other studies, using similar models. (Banerjee et al.,
2002; Wendel, 1981; Krishenbaum et al., 1993)
Myocardial adaptation against oxidative stress is
mediated through augmentation of a number of
cellular antioxidants, such as SOD, catalase,
glutathione peroxidase, glutathione. As IRI is a
common sequel of ischemic heart disease (IHD)
and oxidative stress plays a central role in its
etiopathogenesis, protection against oxidative stress
through a novel mechanism like myocardial
adaptation holds promise as an effective
therapeutic approach.
Myocardial adaptation occurs in response to
various kinds of obnoxious stimuli, like ischemia,
certain endotoxins, reactive oxygen species, etc.
and protects heart from subsequent exposure to
injuries of similar or more severe nature. Although
protective in nature, the basic mechanisms of such
induction of adaptation are harmful in themselves
and, therefore, cannot be relied upon as acceptable
therapeutic methods.
Therefore, a pharmacological, as well as transgenic
approach of myocardial adaptation has recently
become the focus of recent scientific interest. It is
interesting to note that different plants and plant
extracts can also stimulate the synthesis of cellular
antioxidants. It was reported earlier that aged garlic
extract increases the level of SOD, GSH and GPx
in vascular endothelial cell and inhibits oxidative-stress
mediated ischemic-reperfusion damage in rat
brain.
Phenolic compounds, especially flavonoids,
constitute one of the most divers and widespread
group of natural compounds. These compounds
possess a broad spectrum of biological activities
including antioxidant and radical scavenging
properties. (Galvez et al., 2005; Melo et al., 2005;
Parejo et al., 2002) Phenols, a major group of
phytochemicals, have profound importance due to
their AOA.
Protection against ischemia reperfusion induced
oxidative stress in Mimosa pudica plant extract
treated rat hearts was evidenced by preservation of
endogenous antioxidants and prevention in rise of
TBARS. Among the three Mimosa pudica plant
extract treated groups, only 250 mg/kg group
showed the best protection in terms of biochemical
and histopathological evidences. Mimosa pudica
Ph ton 575

5. plant extract in 125 mg/kg group did not show
better histopathological improvement compared to
Mimosa pudica plant extract 500 mg/kg group.
This can be explained by our previous study where
only 250 and 500 mg/kg doses of chronic Mimosa
pudica plant extract administration augmented both
SOD and catalase activities, while 125 mg/kg
Mimosa pudica plant extract administration
augments only SOD activity.
One of the most fashionable ideas for the cause of
reperfusion damage is that the function of cell
membrane is modified by oxygen radicals
generated at the moment of reperfusion (Ferrari et
al., 1991). In one of the study the results also
indicate that the antioxidant activity of E.
officinalis may reside in the tannoids of the fruits of
the plant, which have vitamin C-like properties,
rather than vitamin C itself (Bhattacharya et al.,
1999).
Methionine had no effect on the ventricular or
aortic pressures, heart rate, and myocardial
glutathione levels at any of the time points. In one
of the study showed that methionine has a
significant effect on the myocardial antioxidant
enzyme activities, and only changes in GSHPx
enzyme activity correlated with the mRNA
changes.
These antioxidant changes may have a role in the
beneficial effects of methionine in pathological
rather than physiological conditions (Seneviratne et
al., 1999). Both angiotensin-converting enzyme
inhibitor treatments were associated with increased
nitric oxide production, as assessed by plasma NO-
(3) + NO-(2) level determination, the reports on the
enalapril- and captopril-induced enhancement of
endogenous antioxidant defences and include new
data on glutathione-dependent defences, thus
furthering current knowledge on the association of
ACE inhibition and antioxidants (Cavanagh et al.,
1997).
The ubiquitous presence of some of these stress
genes, such as for HSP 70 and catalase, in normal
unstressed myocardium further suggests a role of
these genes in many basic and essential
biochemical and metabolic pathways. Thus, these
genes are likely to be involved both in the
protection and recovery/repair mechanisms (Das et
al., 1995). The influence of an intake of garlic
powder on the susceptibility to ventricular
arrhythmias under ischemia and reperfusion were
reported in the isolated rat heart (Langendorff
preparation) perfused with a modified Krebs-
Henseleit solution (Isensee et al., 1993).
The chronic garlic intake dose dependently
augmented endogenous antioxidants, which might
have important direct cytoprotective effects on the
heart, especially in the event of oxidant stress
induced injury (Banerjee et al., 2002). Standard
procedure for the estimation of lipid peroxide level
in animal tissues by their reaction with
thiobarbituric acid with external standard,
tetramethoxy-propane was used, and lipid peroxide
level were expressed in terms of nmol
malondialdehyde (Okhawa et al., 1979).
Supplementation of the perfusion medium with
SOD (120 U/ml) and catalase (80 U/ml)
significantly attenuated the I-R injury in S hearts,
and the response in many ways was comparable to
H hearts, hence the therapeutic potential of
increased myocardial endogenous antioxidants
against oxidative stress (Krishenbaum et al., 1993).
Hypoxic preconditioning enhances functional
recovery and reduces cell necrosis following global
ischaemia in the working rat heart, this
phenomenon may, in part, be mediated through
enhanced ascorbate and thiol components of the
antioxidant reserve (Borchgrevink et al., 1989).
Terminalia arjuna Linn (TA) was administered
orally to Wistar albino rats, there was significant
increase in the baseline contents of thiobarbituric
acid reactive substance (TBARS) (a measure of
lipid peroxidation) with both doses of TA.
Significant rise in myocardial TBARS and loss of
SOD, CAT and GSH (suggestive of increased
oxidative stress) occurred in the vehicle-treated
hearts subjected to in vitro IRI. These results
suggested that crude bark of TA augments
endogenous antioxidant compounds of rat heart and
also prevents oxidative stress associated with IRI of
the heart (Gauthaman et al., 2001).
4.1 Outcome of the research
To determine their potential sources in our previous
publications focused on some plants for TPC,
AOA, where in Mimosa pudica (whole plant),
showed a good amount of Total Phenolic Content
(71.23mg/g). IC 50 of Mimosa pudica showed high
(54.13 μg/ml), when they compared with reference
standard Rutin. The DNA nicking activity was
studied on pBR 322 plasmid DNA, the potential
protection was noticed in the Mimosa pudica and
Evolvulus alsinoides L. Since these plant extracts
were the potential sources of natural polyphenols
with promising AOA and a wide range of
biological activities, further research is needed for
the isolation and identification of the active
components can serve as a beneficial source for
herbal therapies in future (Zuhaib et al., 2011).
In this study reported that chronic oral
administration of Mimosa pudica plant extract
caused a significant increase in basal SOD and
catalase activities of rat heart, which was
associated with a concomitant decrease in basal
Ph ton 576

6. lipid peroxidation. Simultaneous increase in SOD
and catalase activities, as observed with chronic
administration of Mimosa pudica plant extract has
special importance of being more beneficial than
increase in SOD activity alone, because without a
concomitant increase in catalase activity, increased
SOD activity may lead to intracellular
accumulation of H2O2 with detrimental effects.
Even though the histopathological profiles of heart
tissue reveals that 500 mg/kg/day for 30 days of
Mimosa pudica plant extract administered rat
hearts subjected to IR injury group showed focal
interstitial edema but no necrosis was observed,
since in this group cytoprotection were observed
with comparative biochemical oxidative enzyme
profiles.
4.2 Future prospects
Cardiac troponin T and troponin I are the most
specific and sensitive laboratory markers of
myocardial cell injury and therefore have replaced
creatine kinase MB as a gold standard ECG and
creatine kinase MB. Identification marker enzyme
CPK, demonstrating the biochemical basis of
protective action of Cl against ISP induced
myocardial injury. Use of PCR and RAPD for
Identification of plasma levels of two markers --
microRNA 122 and 126. The TUNEL assay
identifies single strand DNA breaks with free 3’-
OH terminals. Several studies have raised doubts
about the specificity of TUNEL staining. Collective
evidence suggests that the TUNEL assay is useful
in identifying apoptosis but should be
complemented by additional evidence of apoptosis,
such as the up-regulation of pro- or anti-apoptotic
gene products or structural criterion. In addition,
the anti-apoptotic properties of the herbal extracts
were studied using a combination of techniques of
TUNEL positivity and immunohistochemical
localization of Bax and Bcl-2 proteins.
Conclusion
Hence Mimosa pudica plant extract could be a
therapeutically beneficial for the treatment of
ischemic heart disease (IHD).
Funding and Policy Aspects
This study executed exclusively on self funding.
Author’s Contribution
Sudha kumari. Research Scholar, CMJ University -
- major Research work carried out
Corresponding Author: Dr. Muralidhar T.S.*
Professor & Head, P.G.Department of
Biotechnology – project guidance, data
verification, editing and compilation
Chethan Chowdhary, Kavya. S, Parvathi .C, Post
Graduate Biotechnology Students – extensively
supported for literature search, project work
assistance
Aamir javed: jr. Embryologist – Statistical analysis,
graphical abstract preparation
Umesh. H.R – Senior Lecturer – biochemical
estimations
Dr. Sivakami Sundari P. Assistant Professor, Dept.
of Pharmacognosy – Technical Assistance in
experimental execution
Acknowledgment
The authors are extremely grateful to Dr.
Premchandra Sagar, Vice Chairman, Dayananda
Sagar Institutions and Dr. Krishne Gowda,
Director, Dayananda Sagar College of Biological
Sciences, Bangalore-560078, INDIA, for their
immense guidance and support for this project.
Competing interest
The author’s declares that there is no conflict of
interests regarding the publication of this paper.
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