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Effect of Ethanolic Extraction of Moringa oleifera on Paraoxonase and

Arylesterase enzyme activity in High Fat Diet-induced Obesity in Rats

Article  in  Research Journal of Pharmacy and Technology · October 2018

DOI: 10.5958/0974-360X.2018.00842.9

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 Research J. Pharm. and Tech. 11(10): October 2018

ISSN 0974-3618 (Print) www.rjptonline.org

0974-360X (Online)

RESEARCH ARTICLE

Effect of Ethanolic Extraction of Moringa oleifera on Paraoxonase and

Arylesterase enzyme activity in High Fat Diet-induced Obesity in Rats
Entedhar Rifaat Sarhat, Siham A. Wadi, Ayhan R. Mahmood
Department of Basic Science, Dentistry College, Tikrit University, Iraq
College of Veterinary Medicine , University of Tikrit , Iraq
*Corresponding Author E-mail: entedharr@tu.edu.iq
ABSTRACT:
Background: Obesity is a major health concern both in developed and developing countries. The use of herbal
medicines became the subject of interest for the management of obesity due to its natural origin, cost
effectiveness and minimal side effects. The present study aimed at investigating anti-obesity potential of
Moringa oleifera (MOE) in rats. Forty adult rats (135-160g) was divided into four experimental groups: The first
considered as control negative group (C-ve) and fed on normal diet, while other three groups fed on high fat diet
for three weeks to induce obesity. Obese rats were divided into three equal groups (n= 10 rats). Second group
(obese rats) considered as (C +ve). Third group (obese rat) receivedMOE 600 mg/ kg b.wt. for 12 weeks and
fourth group receivedSimvastatin at a daily dose of 5 mg/ kg b.wt. for 12 weeks, Paraoxonase(PON),
Arylesterase,Total antioxidant status (TAS),malondialdehyde (MDA), superoxide dismutase (SOD) and catalase
(CAT) were evaluated. The results shown significant increase in BMI, and MDA, whereas decrease in (PON,
Arylesterase,TAS, SOD, and CAT) in obese group. Treatment of obese rats with MOE or Simvastatin for 12
weeks produced a significant elevation in PON, Arylesterase, TAS,SOD, and CAT, whereas reduction in body
weight and MDA. Thus, current findings reinforce the advice recommending consumption of Moringa oleifera
to modulate obesity .Conclusion: Conclusively, MOE showed anti-obesity properties by inhibition of PON,
Arylesterase, TAS, SOD, and CAT, and modulation of adipocytes markers.

KEYWORDS: Moringa oleifera, HFD, paraoxonase, Arylesterase.

INTRODUCTION: Consequently, more trials have been conducted on the

Obesity is one of the most common disorders
encountered in clinical practice., because it is a major
risk factor for metabolic syndrome, such as diabetes
mellitus type 2, cardiovascular diseases and several
types of cancer(1,).
use of herbal medicines that were reported to possess
antiobesity potential in-vitro and in-vivo. These herbal
medicines became the subject of interest due to its
natural origin, cost effectiveness and minimal side
effects(2).

There are many ways to prevent or control obesity,
which includes, diet regimes, exercise and medication.
However, the use of anti-obesity drugs such as
Orlistatand Sibutramine has been reported to cause
adverse side effects including high blood pressure,
constipation, dry mouth, headache, heart attack and
insomnia.
Received on 06.09.2017 Modified on 12.10.2017
Accepted on 20.11.2017 © RJPT All right reserved
Research J. Pharm. and Tech 2018; 11(10): 4601-4604
DOI: 10.5958/0974-360X.2018.00842.9
Moringa oleifera (MO) is a member of family
Moringaceae, commonly known as drum stick is a
medicinal plant widely grown in the tropical and
subtropical regions. Moringa oleifera is also known for
its antioxidant activity, essentially due to the presence of
high amounts of polyphenols(3).
Simvastatinthat inhibits the cholesterol synthesis by
inhibiting the enzyme(3-hydroxy-3-methylglutaryl-
coenzyme A) HMG-CoA reductase, is reported to
increase the serum paraoxonase activity(4,5).
Antioxidants via improving the antioxidant defense

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 Research J. Pharm. and Tech. 11(10): October 2018
system offer a promising approach to protect neuronal
cells by removing free radicals, scavenging reactive
oxygen species (ROS) or their precursors, maintaining
redox homeostasis, and decreasing oxidative
damage.The antioxidant defense system can be
enhanced exogenously by ascorbic acid, lipoic acids,
polyphenols, and carotenoids, or endogenously by
catalase, superoxide dismutase (SOD) and glutathione
peroxidase (GPx). Beneficial effects of antioxidants in
reducing oxidative stress in neuronal damage have been
shown in many cell culture models(6).
Many polyphenols in foods are potent antioxidants,and
they have been linked with the hypothesis that their
redox activities may confer specific health benefits(7).
Polyphenols have been shown to act as metal chelators,
preventing the metal-catalyzed formation of radical
species, antioxidant enzyme modulators, and scavengers
of free radicals, including the hydroxyl radical (•OH),
superoxide anion (O2−), nitric oxide (NO), and
peroxynitrite (ONOO−)( 8).
Lipid peroxidation leads to a cascade of reactions,
thereby not only destroys membrane lipids but also
generates endogenous toxicants that can readily react
with adjacent molecules like membrane proteins or
diffuse to more distant molecules like DNA, which may
lead to more hepatic complications and functional
anomalies( 9).
MATERIALS AND METHODS:
Induction of Obesity:
High-fat diet (HFD) induced obesity in rats is
considered to be a reliable tool for the evaluation of
antiobesity activity. HFD that consists of 58% fat, 25%
protein and 17% carbohydrate, lard (13%), cholesterol
(1%), vitamin, and minerals (0.6%) as a percentage of
total kcal ad libitum, respectively, was administered
every(10). Food intake was calculated every day and
bodyweight was measured once in every two days.
Preparation of Moringa oleifera extract:
The air-dried powdered aerial parts of Moringa oleifera
(2 kg) were extracted by cold percolation with 95 %
ethanol (3 x 4 L) till exhaustion. The ethanol extract was
concentrated under reduced pressure to give 250 g of a
brown residue.
Experimental Animal:
The experimental study was carried out on 40 adult rats
(about 135-155 g), during the period from January to
June 2017. The animals were divided into 4 groups each
group consists of 10 animals:
Group 1: healthycontrolrats
Group 2: Obese rats receivedHFD
Group 3: Obese rats receivedM. oleifera 600 mg/ kg
b.wt. for 12 weeks rendering to Jain et al.(2010).
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Group 4: Obese rats receivedSimvastatin at a daily dose
of 5 mg/ kg b.wt. for 12 weeks. Rendering to
Mbikay(11).
Paraoxonase and arylesterase activity of PON1 enzyme
were measured using paraoxone and phenylacetate as
the substrates(12). Malondialdehyde was estimated by the
thiobarbituric acid assay method of terbauer(13).
Superoxide dismutase activities were assayed according
to the procedure described by Woollians(14) and catalase
by Aebi(15). TAS was estimated by the metmyoglobin-
ABTS method( 16).
RESULTS:
The results of the current study revealed that there is
significant increase in BMI (P <0.05) in obese group
with respect to the lean control group (0.341±0.097 VS
0.269±0.097). On the other hand, there is significant
decrease in BMI (P <0.05) of obese groups treated with
MOE or Simvastatin versus the obese control group
(0.308±0.097,or0.295 ±0.097 VS 0.341±0.097).
Results showed that there is significant decrease in
serum PON, arylesterase, TAS,catalase, and SODlevels
(P˂0.05) in obese group when compared with the lean
control group{(149±0.08 vs 203±0.18 U/L),( 170±7.734
vs293±18.08 μmol/L) , (1.8034±0.3349 vs 3.69±0.3834
mmol/l ), (21.661±1.456 vs 27.370±1.534 K/ml), and
(84.22±0.08 vs 112.34±0.07) respectively}. While,
serum MDA level recorded significant increase (P<0.05)
in obese group as compared to the lean control group
(5.801±0.171 vs4.384±0.447).
In contrast, treatment of obese group with the ethanolic
extract of Moringaoliefera or with Simvastatin elicited
significant increase in serum PON, arylesterase,
catalase, and SOD levels (p<0.05) when compared with
those in the obese control group{ (186.54±0.12, or
190.07±±0.1 vs149 ±0.08), ( 257 ±15.70, or 250±17.1 vs
170±7.734), (2.209 ± 0.2183, or 2.8 ± 0.2569 vs 1.8034
± 0.3349mmol/l), (24.340±1.507, or 25.090±1.757 vs
21.661±1.456 K/ml), (104.42±2.89, or109.9±4.55 vs
84.22±3.54 unit/mg x10-1}.
The opposite was observed regarding serum MDA level
which showed significant decrease (p<0.05) in group
treated with Moringa oleifera ethanolic extract or with
Simvastatin in comparison with that recorded in the
obese control group (3.981±0.427, or 4±0.6566, vs
5.801±0.171 mmole/gx 10-2).
 Research J. Pharm. and Tech. 11(10): October 2018

Fig.1: PON level of rats fed a high fat diet and treated with Fig.5: SOD level of rats fed a high fat diet and treated with
Moringa oleifera or simvastatin. Moringa oleifera or simvastatin.

Fig.2: Arylesterase level of rats fed a high fat diet and treated with Fig.6: Catalase level of rats fed a high fat diet and treated with
Moringa oleifera or simvastatin. Moringa oleifera or simvastatin.

DISCUSSION:
Treatment of obese groups with the ethanolic extract of
Moringa oleifera or with the anti- Simvastatin, resulted
in a significant elevation in both PON, and arylesterase,
TAS, CAT, and SOD, whereas reduction in MDA and
BMI in comparison with the obese group control group.
Antioxidants are capable of reducing oxidative stress by
scavenging free radicals. Polyphenols are the most
abundant antioxidants in human diet and are widespread
constituents of fruits and vegetable.
Interestingly, studies have shown that MO leaves are
Fig.3: TAS level of rats fed a high fat diet and treated with
Moringa oleifera or simvastatin.
rich sources of antioxidant due to the presence of
various phytochemicals such as polyphenolics,
carotenoids, α-tocopherol, ascorbic acid, and several
amino acids. The bioavailability of these metabolites in
MO has been directly linked to various biological
profiles, especially its scavenging activity which helps
ameliorate the damaging effects caused by oxidative
stress in diabetic rats(17).
These antioxidant compounds may be responsible for
the increased activity of paraoxonase through direct
interaction with enzyme or stabilization of paraoxonase
on HDL and/or its expression. There are evidences
suggesting that PON1 is an HDL associated antioxidant
Fig.4: MDA level of rats fed a high fat diet and treated with enzyme protecting against atherosclerosis by preventing
Moringa oleifera or simvastatin. from lipoprotein oxidation and hydrolyzing oxidized
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 Research J. Pharm. and Tech. 11(10): October 2018
cholesterol and/or phospholipids in atherosclerotic
lesions and oxidized LDL(18,19). PON1 is readily
inactivated by exogenous or endogenous oxidant
agents(20). Polyphenols may upregulate PON1 expression
and release by sequential activation of protein kinase A
and PPAR-γ(21).Therefore, dietary antioxidants are found
to increase PON1 activity/expression in animals or
humans.
Serum PON1 activity and concentration have also been
shown to be modulated by lifestyle and dietary factors(
22)
. ingestion of polyphenols by apolipoprotein E-
deficient mice (whose plasma PON1 level is lower than
controls), was related with an increase in serum PON
activity(18). The lower TAS observed in test rats than
control rats may indicate the supply of antioxidant by
Moringa Oleifera. Antioxidants like superoxide
dismutase and glutathione peroxidase are jointly
accesses and they are both known to counteract free
radicals (reactive oxygen species). Superoxide
dismutase enhances the breakdown of superoxide anion
into oxygen and hydrogen peroxide. It should be noted
that hydrogen peroxide also is a reactive oxygen species
that can function as a free radical.In the light of this,
catalase or glutathione peroxidase, another known
ntioxidant enzymes has to come in place to convert
hydrogen peroxide into water and oxygen. Moringa
oleifera contains all the antioxidants being accessed by
accessing total antioxidant status because elevation of
any of the components may present with increase total
antioxidant status(23).
CONCLUSION:
MOE elevated the reduced PON, Arylesterase, SOD,
and CAT, whereas lower the elevated MDA. The effects
may be due to the presence of flavonoids in MO, which
helps to speed up the natural healing process in obese
rats. The antiobesity influence of Moringa oleifera
could be attributed to its antioxidant and anti-
inflammatory capacity.Results of this study highlights
the basis for future investigations of MO as a source of
natural product that has the potential to be developed as
medicinal ingredients for prevention and treatment of
obesity and other metabolic diseases in human
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