UTILIZATION OF CLOVE OIL TO REDUCE SOME BIOCHEMICAL ...

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Naglaa et al. World Journal of Pharmacy and Pharmaceutical Sciences

UTILIZATION OF CLOVE OIL TO REDUCE SOME BIOCHEMICAL

DISTURBANCES INDUCED BY HIGH FAT DIET IN RATS

Naglaa E. Mohamed and Mervat M. Anwar

Nuclear Research Center, Atomic Energy Authority, P.O.13759, Egypt.

ABSTRACT

Twenty four male albino rats were divided into four groups. Group 1

(control) fed on a standard diet, group 2 was administered orally clove

oil (200 mg/kg b.wt/day), group 3 fed on high fat diet (HFD) by adding

saturated palm oil to the standard diet (15 g oil /100 g diet) and group 4

was administered orally clove oil + high fat diet for 60 days.

Thiobarbituric acid reactive substance (TBARS), glutathione (GSH)

content and catalase activities (CAT) were estimated in cardiac and

hepatic tissues. Serum lipase, total lipids, phospholipids, cholesterol, triglycerides, lactate

dehydrogenase (LDH), creatinine kinase (CK), Insulin, homeostasis insulin resistance and

plasma glucose were determined. Also, serum free thyroxin (FT4) and free triiodothyronine

(FT3) were estimated. Serum tumor necrosis factor-α (TNF-α) and 8- hydroxyl

deoxyguanosine (8-OHDG) were measured, and histological examination of cardiac and

aortic tissues of rats were observed. The obtained results of rats fed on HFD showed a

significant increase in TBARS and significant decreases in GSH content and CAT activities

in cardiac and hepatic tissues and significant decrease in serum lipase. Significant increases

in serum total lipids, phospholipids, cholesterol, triglycerides, glucose, LDH, CK, insulin and

homeostasis insulin resistance were recorded. Significant decreases in serum FT4 and FT3

were recorded. Significant increases in serum TNF-α and 8-OHDG were recorded. The

histological examination of cardiac and aortic tissues of rats fed on HFD showed some

degenerative cells in theses tissues. Conclusion: It could be concluded that the administration

of clove oil to rats ameliorated the hazardous effects of high-fat diet.

KEYWORDS: High-fat diet, Clove oil, Lipid profile, DNA oxidative stress, Tumor necrosis

factor-α, Thyroid hormones, Insulin.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 9, Issue 4, 50-66 Research Article ISSN 2278 – 4357

*Corresponding Author

[email protected],

Article Received on

27 July 2019,

Accepted on 01 August 2019,

Online on 09 March 2020

DOI: 10.20959/wjpps20204-14565

mailto:[email protected]


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1- INTRODUCTION

Hyperlipidemia is the term used to denote raised serum levels of one or more of total

cholesterol, low-density lipoprotein cholesterol, triglycerides, or both total cholesterol and

triglycerides (combined hyperlipidemia).[1]

The hyperlipidemia can be either primary or

secondary to other diseases but secondary hyperlipidemia is the most common form that can

be induced as a result of endocrine disorders, pancreatitis, cholestasis, protein-losing

nephropathy, obesity and high-fat diets.[2]

The principle metabolic causes of atherosclerosis include hyperlipidemia, hypertension,

obesity, insulin resistance and diabetes mellitus.[3]

The hyperlipidemia is associated with

heart diseases which are the main cause of death.[4]

Atherosclerosis is predominantly a

chronic low-grade inflammatory disease of the vessel wall.[5]

In atherosclerosis, inflammation

starts and evolves in response to cholesterol accumulation in the arterial intima of the large

and medium arteries.[6,5]

Inflammation is reported to promote cardiovascular disease[7]

, which

can be induced by the consumption of a high-fat diet.[7,8]

Some natural compounds are used for improving the cardiovascular disturbances and may

lower the high cholesterol level without using synthetic drugs which have potential side

effects.[9]

The herbal treatment for hyperlipidemia showed no side effects and is relatively

cheap and locally available[4]

while essential oils and plant extracts of aromatic and medicinal

plants are used for such treatment.[10]

Cloves (Syzygium aromaticum L.) are the aromatic dried flower buds of a tree in the family

Myrtaceae and their essential oils are available such as clove bud oil, clove stem oil and clove

leaf oil. Clove bud oil is the best quality which contains eugenol (80-90%) that makes this oil

very potent.[11]

The major constituents of the essential oil of clove are phenylpropanoids such as carvacol,

thymol, eugenol, and cinnaldehyde.[12]

The studied biological activity of clove oil showed

that it has antioxidant[13]

, metal chelation[14]

, neuroprotection[15]

, antibacterial[12]

,

gastroprotection[16]

, renal, cardioprotection[17]

and hepatoprotection properties.[18]

The present study was carried out to evaluate the possible hypolipidemic effect of clove oil

against high-fat diet that induces inflammation, oxidative stress, some biochemical and

histological disorders in male rats.


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2- MATERIALS AND METHODS

2-1- Clove oil

Clove buds were purchased from local market in Egypt. Clove buds were exposed to 5KGy

as preservative tools of food by using Co 60 irradiators at Nuclear Research Centre, Egyptian

Atomic Energy Authority. Then the essential oil of clove buds was obtained by steam

distillation according to the method described in the British Pharmacopeia[19]

by using

Clevenger apparatus for determination of essential oils lighter than water. The volatile oil was

collected after 3 hrs distillation then the oil was filtered and dried over anhydrous sodium

sulfate then the percentage of the oil was calculated as follows: oil % = read of the apparatus /

weight of the herb × 100.[20]

2-2- Palm oil

Saturated palm oil without antioxidant (QC-F-8.2.4-57 Rev ≠ 0) was obtained from Arma

Company, Egypt.

2-3- Experimental animals

Twenty four male albino rats were 7-9 weeks of age weighing 100±10 g were obtained from

the animal house of Nuclear Research Centre, Egypt. The rats were housed in plastic cages in

a well-ventilated room under normal hygienic conditions with 12 hours light - dark cycle

before and during the experiment. The rats were fed on a standard diet and water ad libitum.

The study was carried out at Nuclear Research Centre, Egyptian Atomic Energy Authority.

The manuscript was approved by the scientific committee of publication, committee No.

(158) of the Egyptian Atomic Energy Authority. All rats were handled in accordance with the

standard guide for the care and use of laboratory animals Published by The US National

Institutes of Health (NIH publication No85-23, 1996).

2-4- Experimental design

Rats were divided randomly into four groups each contains 6 rats.

Group 1 (control): rats fed on a standard diet.

Group 2 (clove oil): rats administered an oral dose of clove oil (200 mg/kg b.wt/day) by

gavage using a stomach tube for 60 days.

Group 3 (HFD): rats fed daily for 60 days with a high-fat diet which was prepared by adding

saturated palm oil to the standard diet (15 g oil/100 g diet).

Group 4 (clove oil + HFD): rats fed on a high fat diet and received an oral dose of clove oil

(200 mg/kg b.wt/day) by gavage using a stomach tube for 60 days.


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2-5- Collection of blood

At the end of 60 days, rats were left for 12 h fasting period. Rats were anesthetized then

sacrificed and blood was collected through cardiac puncture. Blood samples were divided

into two parts; one part on sodium fluoride (to prepare plasma) and the other part on the plain

tube (to prepare serum). Blood was centrifuged to collect serum and plasma then stored

frozen at -20°C until biochemical assays.

2-6- Preparation of tissue homogenate

Heart and liver were dissected out, blotted of blood, homogenized separately in phosphate

buffer (pH 7.4) then kept at -20°C for biochemical assays.

2-7- Hormonal and biochemical analysis

Oxidative stress in heart and liver homogenates as thiobarbituric acid reactive

substance(TBARS)was determined according to method of Yoshioka et al.[21]

Glutathion (GSH)content and catalase (CAT) were determined acoording to methods of

Beutler et al[22-23]

respectively.

Plasma glucose, serum lactate dehydrogenase (LDH) and creatinine kinase (CK) were

determined according to methods of Tietz[24,25,26]

respectively.

Serum total lipids, phospholipids, total cholesterol, triglycerides and lipase were determined

according to methods of Zollner and Kirsch,[27-31]

respectively.

Alanine transaminase (ALT), aspartate transaminase (AST), gamma- glutamyl transferase

(GGT) and alkaline phosphatase (ALP) were estimated according to methods of Reitman and

Frankel[32-34]

, respectively.

Serum-free thyroxin (FT4), free triiodothyronine (FT3) and insulin were assayed by

radioimmunoassay (RIA) depending on solid phase RIA using kits obtained from

Immunotech A Beckman Coulter Company. Insulin resistance was calculated according to

the homeostasis model assessment (HOMA-IR).[35]

Serum tumor necrosis factor alpha (TNF-

α) and 8- hydroxyl deoxyguanosine (8-OHDG) were determined using enzyme linked

immune sorbent assay (ELISA), cloud, Clona Cop. USCN, life Science Inc.


54


2-8- Histological examination

For histological examination, pieces of heart and aorta were quickly removed then fixed in

10% neutral buffered formalin solution. The sections were stained with haematoxylin and

eosin then examined under an Olympus light microscope.

2-9- Statistical analysis

The results were presented as the mean ± SE. One way analysis of variance (ANOVA)

followed by Duncan multiple range test was applied for statistical analysis.[36]

3- RESULTS

3-1- Biochemical study

Table (1) showed a significant increase (P<0.05) in TBARS and a significant decrease

(P<0.05) in GSH content and CAT activities of cardiac and hepatic tissues respectively in

group of rats fed on HFD. However, the administration of clove oil improved these effects as

compared to the control group by decreasing TBARS, and increasing GSH content and CAT

activities in cardiac and hepatic tissues respectively. GSH content and CAT activities in

cardiac and hepatic tissues.

The results of this study revealed that rats fed on HFD showed a significant increase (P<0.05)

in serum total lipids, phospholipids, total cholesterol, and triglycerides, as well as a

significant decrease (P<0.05) in lipase as compared to control group. Significant decreases

(P<0.05) in FT3 and FT4 in rats fed on HFD were determined as compared to control group.

While rats administered clove oil + HFD showed improvement in these parameters (Table 2).

The obtained results showed significant increases (P<0.05) in AST, ALT, GGT, ALP, LDH,

and CK in rats fed on HFD as compared to control group. While rats fed on clove oil + HFD

showed improvement in these parameters (Table 3).

The results showed significant increases (P<0.05) in glucose, insulin, and insulin resistance

index (HOMA-IR) in rats fed on HFD as compared to control group. While rats administered

clove oil + HFD showed improvement in these parameters (Table 4).

Data in the table (5) showed that rats fed on HFD revealed a significant increase (P<0.05) in

TNF-α, and 8-OHDG as compared to the control group. While rats fed on clove oil +HFD

revealed significant improvement in these parameters.


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Table (1): Effect of clove oil and/or high fat diet (HFD) on TBARS level, GSH content

and CAT activities in cardiac and hepatic tissues of different rat groups.

Organs

Groups

Control Clove oil HFD Clove oil +

HFD

Heart

TBARS (nmol/g wet tissue)

83.66±3.88c

81.55±6.05c

137.02±7.17a

99.59±3.63b

GSH (mg/g wet tissue) 25.70±0.57a 26.30±0.98

a 13.71±0.30

b 26.73±0.81

a

CAT (U/g wet tissue) 108.65±3.36a 109.29±4.09

a 75.75±3.04

c 95.19±2.87

b

Liver

TBARS (nmol/g wet tissue) 44.07±4.57

b 43.92±3.03

b 129.02±9.25

a 99.27±4.36

b

GSH (mg/g wet tissue) 36.32±1.19a 35.75±0.72

a 23.66±0.21

c 29.21±0.64

b

CAT (U/g wet tissue) 192.22±1.78a 190.34±3.46

a 124.82±4.05

b 183.34±7.23

a

All values are means ± SE.

Values with different superscripts in the same rows are significantly different (P<0.05).

Table (2): Effect of clove oil and/or high fat diet (HFD) on serum total lipids,

phospholipids, cholesterol, triglycerides, lipase, FT3 and FT4 of different rat groups.

Parameters Groups

Control Clove oil HFD Clove oil + HFD

Total lipids (mg/dL) 702.09±14.32c 719.12±13.92

c 860.45±23.55

a 771.68±15.76

b

Phospholipids (mg/dL) 163.58±1.45c 154.76±4.65

c 223.88±5.45

a 170.14±2.30

b

Cholesterol (mg/dL) 104.14±6.12c 105.13±6.11

c 262.31±12.68

a 185.42±4.34

b

Triglycerides (mg/dL) 97.93±1.95c 100.68±2.58

c 131.78±4.73

a 109.77±2.21

b

Lipase (U/L) 19.07±2.26a 19.76±2.28

a 7.58±2.12

c 13.17±0.92

b

FT3 (pg/mL) 2.08±0.21b 2.00±0.21

b 1.79±0.06

a 1.90±0.20

b

FT4 (ng/dL) 2.19±0.23a 2.22±0.24

a 1.60±0.13

b 2.00±0.16

a



Table (3): Effect of clove oil and/or high fat diet (HFD) on ALT, AST, GGT, ALP, CK

and LDH of different rat groups.

Parameters Groups


ALT (U/L) 65.17±5.59c 60.83±4.83

c 127.17±8.85

a 80.5±3.97

b

AST (U/L) 73.4±3.82c 74.16±3.18

c 114.17±1.22

a 86.66±1.35

b

GGT (U/L) 17.68±0.81b 18.78±4.53

b 32.78±2.96

a 19.06±1.57

b

ALP (IU/L) 84.91±4.99c 81.09±5.96

c 101.16±3.50

a 79.54±3.47

b

CK (U/L) 145.54±6.71c 148.68±6.61

c 459.95±9.77

a 354.41±3.99

b

LDH (U/L) 457.64±36.94c 455.2 1±26.37

c 946.44±27.30

a 750.00±42.52

b

All values are means ±SE.



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Table (4): Effect of clove oil and/or high fat diet (HFD) on insulin, glucose and insulin

resistance (HOMA-IR) of different rat groups.

Parameters Groups


Insulin (ng/L) 12.19±0.53b 12.96±0.87

b 21.27±0.72

a 15.50±0.86

b

Glucose (mg/dL) 71.18±9.11b 73.77±10.39

b 123.88±7.73

a 79.11±9.94

b

HOMA-IR 8.61±0.94b 10.19±0.86

b 13.66±0.71

a 9.91±0.68

b



Table (5): Effect of clove oil and/or high fat diet (HFD) on serum tumor necrosis factor

– alpha (TNF-α) and 8- hydroxyl deoxyguanosine (8-OHDG) of different rat groups.

Parameters Groups


TNF- α (pg/mL) 190.41±3.88c 188.25±4.70

c 260.93±3.17

a 215.70±3.25

b

8-OHDG(ng/ mL) 4.98±0.32c 4.14±0.39

c 18.76±0.61

a 10.02±0.31

b

All values are means ± SE.,


3-2- Histological study

The histological changes related to clove oil + HFD were observed in cardiac (figure 1) and

aortic (figure 2) tissues. The examination of normal cardiac tissues showed normal

histological structure (fig. 1a). On the other hand, sections of cardiac tissues of rat fed on

HFD (fig. 1b) showed intermuscular hemorrhage, haemosiderosis, vacuolations in the wall of

blood vessel, perivascular edema, dilatation and necrosis of cardiac myocytes associated with

inflammatory cells and congestion of myocardial blood vessels. On the other hand, sections

of rats administered clove oil + HFD (fig. 1c) showed intermuscular hemorrhage.

The histological examination of aortic tissues revealed no histological changes in the control

group (fig. 2a). The section of the aorta of rat fed on HFD showed vacuolations and focal

necrosis of tunica media (fig. 2b) while sections of the aorta of rat administered clove oil +

HFD showed slight vacuolations of tunica media (fig. 2c).


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Figure (1): Photomicrographs of rat’s cardiac sections (H&E X400):

Fig. (1a): control group, fig. (1b): HFD group and fig. (1c): clove oil + HFD group.

Figure (2): Photomicrographs of rat’s aortic sections (H&E X400):

Fig. (2a): control group, Fig. (2b): HFD group and fig. (2c): clove oil + HFD group.


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4- DISCUSSION

Hyperlipidemia is the result of oxidative stress due to the interaction of HFD with free

radicals. The elevated level of TBARS in hepatic and cardiac tissues of rats fed on a high fat

diet is a clear manifestation of excessive formation of free radicals and activation of lipid

peroxidation. Furthermore, the increment in the levels of total cholesterol and triglycerides of

high fat diet group may be in the levels of serum cholesterol and triglycerides of

hyperlipidemic rats may be a result of lipid peroxidation induced by high- fat diet. This result

is in agreement with Shyamala et al.[37]

who reported that increases of lipid peroxidation level

are associated with increased serum cholesterol and triglycerides levels of hyperlipidemic

rats.

In HFD showed a significant decrease in GSH content and CAT activities in cardiac and

hepatic tissues this depletion was improved in rats in rats fed on clove oil + HFD. The

increase of CAT activities in rats fed on high-fat diet and administered clove oil indicated the

ability of clove oil to decrease oxidative stress induced by clove oil to stop the oxidative

damage induced by hyperlipidemia. The present results are in agreement with Said[38]

who

reported that eugenol can decrease lipid and oxidative stress as well as improving the

intracellular antioxidant defense. The previous study showed that 5 and 10 KGy irradiated

clove buds found that the dose of 5kGy increased the antioxidant activity of clove essential

oil. The antioxidant activity of clove oil can be induced due to its eugenol (77%), eugenol

acetate (15%), β-caryophyllene (2%) and farnesene (4%) which are the main constituents of

such oil.[20]

While other study showed that the essential clove oil was found to be five- fold

higher than that observed for α-tocopherol.[39]

The antioxidant compounds of commercial

clove oil are eugenol and β-caryophyllene; the dominated components that have the ability

for preventing lipid peroxidation.[40]

The present study demonstrated that HFD induced significant increases in serum total lipids,

phospholipids, cholesterol and triglycerides, and a significant decrease in lipase. These results

are in agreement with Thounaojam et al.[41]

The free radicals can be the major reason for hormonal imbalance which induces

hyperlipidemia through its multiple effects on lipid metabolism including increased synthesis

of cholesterol and triglycerides.[42]

Cholesterol is derived from the exogenous diet and

endogenously from acetyl CoA in a series of biosynthesis reactions.[43]

The

hypercholesterolemia is attributed to the increased activation of 3-hydroxyl-3-methyl glutaryl


59


Co-enzyme A (HMG CoA) reductase and the key regulatory enzyme in the reduction of the

overall process of cholesterol synthesis.[44]

Administration of clove oil has been reported to

reduce increased total lipids, phospholipids, Triglycerides and total cholesterol in serum due

to the presence of the active ingredient of clove oil eugenol. The present results showed a

significant decrease in serum total lipids, phospholipids, cholesterol and triglycerides, and

significant increase in lipase in rats fed on HFD + clove oil which is in agreement with

Younies,[45]

These results may be due to eugenol of clove oil. Clove oil might lower the

cholesterol and triglycerides levels then simultaneously reduced the hepatic fatty acid

oxidation. These changes were attributed to the suppression in hepatic fatty acid synthase,

glucose-6-phosphate dehydrogenase, and also may be due to the decreased hepatic-3-

hydroxyl-3-methyl glutaryl Co-enzyme A (HMG CoA) reductase, acyl Co A and cholesterol

acyl transferase.[45]

This-improvement due to phyto compounds (like saponins) in clove oil by

inhibiting the pancreatic lipase activity in mice fed on HFD leading to increased fat excretion

due to reduced intestinal absorption of dietary fats.[46]

The significant decrease in FT3 and

FT4 levels in rats fed on HFD may be due to hormonal disturbance as results of increased

oxidative stress.

The present results showed significant increases in AST, ALT, ALP, and GGT in rats fed on

HFD which are in agreement with.[47,48]

The protective effect of clove oil on HFD induced-

hepatotoxicity in rats was appeared to be related to inhibition of lipid peroxidation in addition

to free radicals scavenging action. The observed antioxidant and hepatoprotective activities of

clove oil may be due to the presence of polyphenolic compounds and flavonoids.[49]

The

present results are in agreement with Abozid and El-Sayed.[50]

The improvement in AST,

ALT, ALP, and GGT in rats fed on high-fat diet + clove oil revealed the hepatoprotective

effect of clove oil.

In the present study, the significant increases in serum CK and LDH in rats fed on HFD

indicated vascular dysfunction which is in agreement with Garjani et al.[51]

who reported that

hypercholesterolemia, even without atherosclerotic lesions, can cause vascular dysfunction.

The rats administered clove oil + HFD revealed significant improvement in CK and LDH

which may be due to the presence of phenolic, polyphenolic, terpenoid and carotenoid

compounds.[52]

The present results indicated that rats fed on HFD showed significantly higher fasting

glucose, insulin, and insulin resistance as compared to the control group. These results are in


60


agreement with other data which showed that rats fed on HFD had developed metabolic

disturbance or type 2 diabetes.[53]

High fat diet can induce increased insulin resistance and

reduce insulin response while insulin resistance induced by an acute increase in plasma free

fatty acids is not rapidly reversible.[54]

The generation of reactive oxygen intermediate has a role in the damage of the cellular

membranes and DNA.[55]

It has been demonstrated that a fatty liver is an insulin resistant as

in the static phase of the living organism, which resulted in elevated glucose and very low-

density lipoprotein production.[56]

The insulin resistance has an important role in the

development of atherogenic dyslipidemia and cardiovascular disease.[10]

Insulin resistance (IR) is considered the key mechanism of obesity, diabetes and heart

diseases.[57]

In the course of months or years, IR is followed by an increase in β-cell insulin

secretion and by several complications known as the insulin resistance syndrome which is

associated with dyslipidemia, hypertension, hyperglycemia and cardiovascular disease.[58]

Rats fed on clove oil + HFD showed improvement in these parameters. This result may be

due to the percentage of eugenol 77% of clove oil.

The present results showed a significant increase in proinflammatory marker (TNF-α) in rats

fed on HFD. This result is in agreement with Sharma et al.[59,60]

The previous study indicated

that TNF-α has been implicated in the pathogenesis of chronic systemic inflammatory

conditions and in the development of atherogenesis and vascular inflammation.[61]

TNF-α can

induce an impairment of endothelium development vasodilation in a variety of vascular beds

by increasing oxidative stress and decreasing the release of Nitric oxide (NO).[62]

The present

results indicated that administration of clove oil with HFD to rats attenuated HFD induced

increases in inflammatory marker (TNF-α).

The present study revealed that hyperlipidaemia (induced by high fat diet) causes a

significant increase in 8-OHDG which indicate increased oxidative stress. However, 8-

OHDG is one of the major reactive oxygen species induced DNA base modified products that

is a sensitive marker of oxidative DNA damage.[63]

This result is in agreement with

Maciejczyk et al.[64,65]

One of the reasons for the occurrence of oxidative DNA damage is free

radicals attack the deoxyribose, which usually leads to breaking a single strand of DNA.[65]

The present results indicated that administration of clove oil with HFD to rats attenuated

HFD induced increases in 8-OHDG due to the percentage of eugenol 77% of clove oil.


61


In the present study, hypercholesterolemia, even without atherosclerotic lesions, can cause

vascular dysfunction as showed in the histological investigations of heart and aorta in rats fed

on HFD. This result is in agreement with Garjani et al.[51]

The present study indicated that clove oil has antioxidant, anti- inflammatory and

hypolipidemic effects against high-fat diet induced-cell damages due to the effect of active

compounds found in the oil. In addition to the improvement in the histological structure of

cardiac and aortic tissues in HFD + clove oil treated rats, the eugenol found in the oil has

potential health benefit toward different metabolic syndromes such as dyslipidemia.

5- CONCLUSION

It could be concluded that the hypolipidemic, hepatoprotective, anti-inflammatory and the

antioxidant effects of clove oil has the ability to reduce the oxidative stress by scavenging

free radicals generated in the body as a result of high-fat diet. In view of the obtained results,

we must change our sedentary life style and avoid high fat diet or fast food, which is

characterized by reduced dietary fiber, fruit and vegetable consumption and increased fat and

refined sugar. Also, our diet must contain naturally antioxidant like clove oil.

Conflict of Interest

The authors declare no conflict of interest. The authors alone are responsible for the content

and writing of the manuscript.

ACKNOWLEDGEMENT

The authors wish to thank Prof. Dr. K.A. Ahmed, Professor of Veterinary Pathology, Faculty

of Veterinary Medicine, Cairo University, Egypt, for performing the histological

examinations.

6-REFERENCES

1. Sivaiah K, Reddy GAK. 2012. Evaluation of anti-hyperlipidemic activity of hydro

alcoholic extract of Moringa oleifera seeds in high fat diet induced rat model. Int J

Pharmacol Scr Methods, 2(2): 72-76.

2. Ahmed HH, Abdalla MS, Eskande EF, Al-Khadragy MF, Massoud MN. 2011. Caulepra

prolifera ameliorates the impact of dyslipidemia induced oxidative stress and

inflammation. Res., 3(2): 110-119.


62


3. Lioyd Jones DM, Hong Y, Labartha D et al. 2010. Defining and setting national goals for

cardiovascular health promotion and disease reduction. The American Heart

Association´s Strategic Impact goal through 2020 and beyond. Circulation, 121(4):

586-613.

4. Kaur G, Meena C. 2013. Evaluation of anti-hyperlipidemic potential of combinatorial

extract of curcumin, piperine and quercetin in triton induced hyperlipidemia in rats. Sci

Int., 1(3): 57-63.

5. Lorenzatti AJ, Retzlaff BM. 2016. Unmet needs in the management of atherosclerotic

cardiovascular disease: is there a role for emerging anti—inflammatory interventions? Int

J Card., 221: 581-586.

6. Gimbrone MA, Garcia-Cardena G. 2016. Endothelial cell dysfunction and the

pathobiology of atherosclerosis. Circ. Res., 118: 620-636.

7. Lumeng CN, Saltiel AR. 2011. Inflammatory links between obesity and metabolic

disease. J Clin Invest, 121(6): 2111-2117.

8. Chalkiadaki A, Guarente L. 2012. High fat diet triggers inflammation induced cleavage of

SIRT1 in adipose tissue to promote metabolic dysfunction. Cell Metab., 16(2): 180-188.

9. Sharma N, Sharma P, Jasuja ND, Joshi SC. 2013. Hypocholesterolemic and antioxidant

potentials of some plants and herbs: a review. RRJZS, 1(2): 26-42.

10. Al-Okbi SY, Mohamed DA, Hamed TE, Edris AE. 2014. Protective effect of clove oil

and eugenol microemulsions on fatty liver and dyslipidemia as components of metabolic

syndrome. J Med Food, 17(7): 764-771.

11. Stichi FD, Smith RM. 2010. Eugenol: some pharmacologic observation. J Dental Res.,

50(6): 1531-1535.

12. Chaieb K, Hajlaoui H, Zmanta T, Kahla-Nakbi AB, Rouabbia M et al. 2007. The

chemical composition and biological activity of clove essential oil, Eugenia caryophyllata

(Syzigium aromaticum L. Myrtaceae) a short review. Phytother Res., 21(6): 501-506.

13. Misharina TA, Samusenko AL. 2008. Antioxidant properties of essential oils from lemon,

grapefruit, coriander, clove and their mixtures. Prikl Biokhim Mikrobiol, 44(4): 482-486.

14. Yadav AS, Bhatngar D. 2007. Modulatory effect of spice extracts on iron induced lipid

peroxidation in rat liver. Biofactors, 29(2-3): 147-157.

15. Nangle MR, Gibson TM, Cotter MA, Cameron NE. 2006. Effects of eugenol on nerve

and vascular dysfunction in streptozotocin diabetic rats. Planta Med., 72(6): 494-500.


63


16. Santin JR, Lemos M, Klein-Junior LC, Machado ID, Costa P et al. 2011. Gastroprotective

activity of essential oil of the Syzygium aromaticum and its major component eugenol in

different animal models. Naunyn Schmiedebergs Arch Pharmacol, 383(2): 149-158.

17. Bafana PA, Balaraman R.2005. antioxidant activity of DHC-1, an herbal formulation, in

experimentally induced cardiac and renal damage. Phytother Res., 19(3): 216-221.

18. Abdel-Waha M A , Ali SE. 2005. Antioxidant property of Nigella sativa (black cumin)

and Syzgium aromaticum (clove) in rats during aflatoxicosis. J Appl Toxicol, 25(3):

218-223.

19. British Pharmacopeia 1988. HMSO, London, 2: 137-138.

20. Anwar MM, Ali S, Rashwan O A. 2010. Antioxidant and antimicrobial activities of

essential oils extracted from gamma irradiated clove and ginger. Isotope and Rad Res.,

42(4): 1007-1024.

21. Yoshioka T, Kawada K, Shimada T, Mori M. 1979. Lipid peroxidation in maternal and

cord blood and protective mechanism against activated oxygen toxicity in the blood. Am

J Obstet Gynecol, 135(3): 372-376.

22. Beutler E, Duran O, Kelly B. 1963. Improved method of blood glutathione. J Lab Clin

Med., 61(51): 882-888.

23. Sinha AK. 1972. Colorimetric assay of catalase. Anal Biochem, 47: 389-394.

24. Tietz NW. 1986. Textbook of clinical chemistry. WB Saunders London Philadelphia,

P.796.

25. Tietz NW. 2005. Textbook of clinical chemistry and molecular diagnostics. 4th

ed. Burtis

CA, Ashwood ER and Bruns DE. WB Saunders Co.

26. Young DS. 2001. Effects of disease on clinical lab. Tests. 4th

ed. AACC. Washington DC.

27. Zollner N, Kirsch K. 1962. Colorimetric method for determination of total lipids. Z Ges

Exp Med., 135: 545-550.

28. Nie Y, He JL, Hsia SL.1993. A micro enzymatic method for determination of choline

containing phospholipids in serum and high density lipoproteins. Lipids, 28(10): 949-951.

29. Allain CC, Poon LS, Chan CS, Richmond W, Fu PCC.1974. Enzymatic determination of

total serum cholesterol. Clin Chem., 20(4): 470-475.

30. Fossati P, Principe L.1982. Serum triglycerides determined colorimetrically with an

enzyme that produces hydrogen peroxide. Clin Chem., 28(10): 2077-2080.

31. Moss DW, Henderson AR. 1999. eds in Tietz textbook of clinical chemistry, 3rd

ed,

Philadelphia, WB Saunders company, P. 689-708.


64


32. Reitman S, Franke SA. 1957. A colorimetric method for the determination of serum

glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol, 28(1): 56-63.

33. Szasz, G. 1969. A kinetic photometric method for serum gamma glutamyl transpeptidase.

Clin Chem., 15(2): 124-136.

34. Belfield A, Goldberg DM. 1971. Revised assay for serum phenyl phosphatase activity

using 4-aminoantipyrine. Enzyme, 12(5): 561-573.

35. Matthews DR, Hosker JP, Rudenski AS, Naylor BA, Treacher DF, Turner, RC. 1985.

Homeostasis model assessment: insulin resistance and beta-cell function from fasting

plasma glucose and insulin concentrations in man. Diabetologia, 28(7): 412-419.

36. Duncan DB. 1955. Multiple range and multiple F-test. Biometrics, 11: 1-42.

37. Shyamala MP, Venukumar MR, Latha MS. 2003. Antioxidant potential of the Syzygium

aromaticum (gaertn.) Linn (cloves) in rats fed with high fat diet. Ind J Pharmacol, 35(2):

99-103.

38. Said MM. 2011. The protective effect of eugenol against gentamicin induced

nephrotoxicity and oxidative damage in rat kidney. Fundam Clin Pharmacol, 25(6):

708-716.

39. Nagabau E, Rifkind M, Boindala S, Nakka L. 2010. Assessment of antioxidant activity of

eugenol in vitro and in vivo. Methods Mol Biol., 610: 165-180.

40. Miguel MG. 2010. Antioxidant and anti-inflammatory activities of essential oils: a short

review. Molecules, 15(12): 9252-928.

41. Thounaojam MC, Jadeja RN, Devkar R, Ramachandran AV. 2009. Dysregulation of lipid

and cholesterol metabolism in high fat diet fed hyperlipidemic rats: protective effect of

sida rhomboidea, roxb leaf extract. J Heal Sci., 55(3): 413-20.

42. Bowden DA, McLean P, Steinmetz A, Fontana D, Mattys C, Warnick GR. et al. 1989.

Lipoprotein apolipoprotein and lipolytic enzyme changes following estrogen

administration in post menopausal women. J Lipid Res., 30(12): 1895-1906.

43. Garcia M, Bayon D, Culebras F, Jorquer P, Garcia D. 1996. Hepatic metabolism of

cholesterol. Nutr Hosp., 11(1): 37-42.

44. Bok S, Lee S, Park Y, Bae K, Son K, Joong T et al. 1999. Plasma and hepatic cholesterol

and hepatic activities of 3-methyl-glutaryl-Co A reductase and acyl Co A, cholesterol

transferase are lower in rats fed citrus peel extracts or a mixture of citrus bioflavonids. J

Nutr., 129: 1182-1185.

45. Younies BM. 2008. Effect of the aqueous extract of cloves (Syzygium aromaticum) on

hyperlipidemia in senile rats. J Rad Res Appl Sci., 1(1): 53-63.


65


46. Han L K, Zheng YN, Xu B J, Okuda H, Kimura Y. 2002. Saponins from platycodi radix

ameliorate high diet induced obesity in mice. J Nutr., 132(8): 2241-2245.

47. Helal EGE, Eid FA, El-Wahsh AMSE. 2011. Effect of fennel (Foeniculum vulgare) on

hyperlipidemic rats. Egy J Hos Med., 43(1): 212-225.

48. Imafidon K E, Okunrobo L O. 2012. Study on biochemical indices of liver function tests

of albino rats supplemented with three sources of vegetable oils. Nig J Basic Appl Sci.,

19(2): 105-110.

49. Palanivel MG, Rajkapoor B, Kumar RS, Einstein JW et al. 2008. Hepatoprotective and

antioxidant effect of Pisonia aculeate L. against CCl4 induced hepatic damage in rats. SCi

Pharm, 76: 203-215.

50. Abozid M M, El-Sayed SM. 2013. Antioxidant and protective effect of clove extracts and

clove essential oil on hyderogen peroxide treated rats. Int J Chem Tech Res., 5(4):

1477-1485.

51. Garjani A, Azarmiy Y, Zakheri A, Akbari N A, Ghosh P, Bitsanis D et al. 2011.

Abnormal aortic fatty acid composition and small artery function in offspring of rats fed a

high fat diet in pregnancy. J Physiol, 533(pt3): 815-822.

52. Henning SM, ZhangY, Seeram N P, Lee R P, Wang P, Bowerman S et al. 2011.

Antioxidant capacity and phytochemical content of herbs and spices in dry, fresh and

blended herb paste form. Int J Food Sci Nutr., 62(3): 219-225.

53. Rinnankoski-Tuikka R, Silvennoinen M, Torvinen S, Hulmi JJ, Lehti M, Kivelä R et al.

2012. Effect of high fat diet and physical activity on pyruvate dehydrogenase kinase 4 in

mouse skeletal muscle. Nutr Metab, 9: 53-65.

54. Han DH, Hancock C, Jung SR, Holloszy JO. 2009. Is fat induced muscle insulin

resistance rapidly reversible? Am J Physiol Endocrinol Metab., 297(1): E236-E241.

55. Edmison J, Mccullough J. 2007. Pathogenesis of non-alcoholic steatohepatitis: human

data. Clin Liver Dis., 11(1): 75-104.

56. Vozarova B, Stefan N, Lindsay RS, Saremi A, Pratley RE, Bogardus C. et al. 2002. High

alanine aminotransferase associated with decreased hepatic insulin sensitivity and

predicts the development of type 2 diabetes. Diab., 51(6): 1889-1895.

57. Bray GA. 2004. Medical consequences of obesity. J. Clin. Endocr. Metab., 89(6):

2583-2589.

58. Reaven G, Abbasi F, McLaughlin T. 2004. Obesity, insulin resistance and cardiovascular

disease. Recent Prog Horm Res., 59: 207-223.


66


59. Sharma H, Joshi A, Lad H, Bhatnagar D. 2018. Anti-oxidative, anti-inflamatory and anti-

atherosclerotic effect of taurine on hypercholesterolemia induced atherosclerotic rats. Int

J Pharm pharmaceut Sci., 10(3): 145-150.

60. Yida Z, Imam MU, Ismail M, Ismail N, Ideris A. 2015. High fat diet induced

inflammation and oxidative stress are attenuated by n-acetylneuraminic acid in rats. J

Biom Sci., 22: 96-105.

61. Ito TK, Yokoyama M, Yoshida Y, Nojima A, Kassai H, Oishi K et al. 2014. A crucial

role for CDC42 in senescence associated inflammation and atherosclerosis. Plos ONE,

9(7): e102186.

62. Zhang H, Park Y, Wu J, Chen XP, Lee S, Yang J, Dellsperger KC, Zhang C. 2009. Role

of TNF-α in vascular dysfunctuion. Clin. Sci (Lond)., 116(3): 219-230.

63. Cakir S. 2015. The investigation of effects on DNA damage in blood of high fat diet and

acrylamide in the rats. Conference: BJCL XXII, A1: Bosnia.

64. Maciejczyk M, Zebrowska E, Zalewska A, Chabowski A. 2018. Redox balance,

antioxidant defense, and oxidative damage in the hypothalamus and cerebral cortex of

rats with high fat diet induced insulin resistance. Oxid Med Cell Long. 6940515: 1-11.

65. Yener Y, Yerlikaya FH. 2018. Western diet induces endogen oxidative deoxy ribonucleic

acid damage and inflammation in Wistar rats. Rev. Nutr., 31(3): 263-273.

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