Effect of Ethanolic extract of Solanum lycopersicum & Vitis vinifera on lipid
profile of Diet and drug induced obesity
Goyal, S.*, Kumar, S.
Research Scholar, School of pharmacy & medical sciences
Singhania university, pacheri bari, jhunjhunu (raj.), india
Address For Correspondence
Shubham Goyal
[email protected]
Introduction
Obesity is a crippling, stigmatized, costly and life shortening disease that has no single
metabolic origin. According to the World Health Organization (WHO), obesity is increasing at
alarming rates in industrialized and ‘industrializing’ world and is considered to be a disorder of
energy balance [1]. Orlistat (inhibitor of pancreatic and gastrointestinal lipases), prevents the
absorption of approximately 30% of dietary fat, but is associated with side-effects like renal
failure, oily spotting, liquid stools, fecal urgency or incontinence, flatulence and abdominal
cramping [2] and Sibutramine (inhibits norepinephrine and serotonin neuronal uptake) but is
associated with side-effects like insomnia, dry mouth, increases in blood pressure, heart rate,
cardiac depression [2].
Solanum lycopersicum is known to have lipid-lowering effects and antioxidant activities.
Red tomato paste decreased the serum levels of total cholesterol and low-density lipoprotein
(LDL) cholesterol and increased the high-density lipoprotein (HDL) cholesterol levels in
hamsters fed a high-cholesterol diet. Solanum lycopersicum is a antioxidant and also have an
thermogenic effect However, and little is known about the effects of Solanum lycopersicum on
diet-induced and antipsychotic drug induced obesity . Vitis vinifera is one of the most commonly
consumed fruits in the world. It has various biological functions, due to its rich polyphenol
ingredients, most of which are contained in its seeds (60-70%) and skin (30%). However, large
quantities of grape seed wastes are produced annually by the food processing industry-wine,
juice etc [3]. Vitis vinifera ingredients also have healthful properties. Phenolic bioflavanoids
from grape seeds and skins are powerful antioxidants. When used with either vitamins C or E,
the extract possesses double the antioxidant capacity [4]. Polyphenol in Vitis vinifera seeds have
also been reported to have a variety of biological activities, including antioxidant,
antithrombotic, and cardioprotective effects [5]. However, little is known about the effects of
Vitis vinifera extracts on diet-induced and antipsychotic drug induced obesity.
Material & Methods
High fat diet-induced obesity
Experimental obesity was produced by feeding high fat diet [6] to rats for a period of 8
weeks.
Antipsychotic drug (sulpiride) induced obesity
Experimental antipsychotic drug induced obesity was produced by administration of
Sulpiride (20 mg/kg i.p.) for 4 weeks [7].
The experimental protocol was approved by the Institutional Animal Ethical Committee (IAEC)
via letter no IEC/010/23-02
Plant material
Solanum lycopersicum fruits were collected in the months of January–February 2011
from New grain market-Sector 26, Chandigarh, India, authenticated in the Botany Department
and a voucher spec-imen (No. Pharmacy/SLF/ 08-09/01/SG) was deposited in the Pharmacy
Department of the university.Vitis vinifera Fruits were collected in the months of March–April
2011 from New grain market-Sector 26, Chandigarh, India,, authenticated in the Botany
Department and a voucher spec-imen (No. Pharmacy/VNF/ 08-09/02/SG) was deposited in the
Pharmacy Department of the university.
Preparation of Ethanolic extract from Solanum lycopersicum Fruits
Solanum lycopersicum was carefully washed; skin and seeds were manually separated
from the pulp. Then the pulp was homogenized in a blender (Philips 2200) and extracted three
times with MeOH (3 × 15 L each) at room temperature in the dark for 12 h per extraction. The
mixture was sonicated (Buchi-R-215, Germany) for 5min and then filtered through gauze twice.
The filtrate was evaporated under vacuo (Buchi-R-215, Germany) to remove methanol.
Preparation of Ethanolic extract from Vitis vinifera Fruits
The fruits was homogenized in a blender (Philps 2200) and extracted three times with
ethanol (3X1.5 L) in a soxhlet appa-ratus. The filtrates were then combined and filtered and
concentrated to dryness in a rotary evaporator (Buchi-R-215, Germany) to obtain a crude
ethanolic extract.
Assessment of biochemical parameters
1. Estimation of serum biochemical parameters
The total cholesterol was estimated by cholesterol oxidase peroxidase CHOD-POD method
[8], serum triglyceride was estimated by glycerophosphate oxidase peroxidase GOD-POD
method [9], HDL was estimated by cholesterol oxidase peroxidase CHOD-POD method [8],
LDL concentrations were calculated from the Friedewald equation [10] using commercially
available kit (Medsource Ozone Biomedicals Pvt. Ltd. Faridabad, India).
2. Tissue Biochemical Estimations:
Assessment of Renal Oxidative Stress
The development of oxidative stress in the kidney was assessed by estimating tissue
thiobarbituric acid reactive substances (TBARS) [11] and reduced glutathione content [12].
Experimental protocol
Thirteen groups of wistar albino male rats were employed in the present study. Each group was
comprised of six animals. All animals are randomly divided into different groups.
Group I: Normal Rats were maintained on standard chow diet and water ad libitum.
No treatment was given to these rats.
Group II: Normal rats were maintained on high fat diet for six weeks to produce obesity.
Group III: Sibutramine (10mg/kg/day p.o., 8 weeks) was administered to rats on High fat diet
and continued upto the end of the eighth week.
Group IV: Vitis vinifera (250mg/kg/day p.o., 8 weeks) was administered to rats on High fat diet
and continued upto the end of the eighth week.
Group V: Vitis vinifera (500mg/kg/day p.o., 8 weeks) was administered to rats on High fat diet
and continued upto the end of the eighth week.
Group VI: Solanum lycopersicum (250mg/kg/day p.o., 8 weeks) was administered to rats on
High fat diet and continued upto the end of the eighth week.
GroupVII: Solanum lycopersicum (500mg/kg/day p.o., 8 weeks) was administered to rats on
High fat diet and continued upto the end of the eighth week.
Group VIII: Sulpride (20mg/kg/day i.p, 4 weeks) was administered to Female rats.
Group IX: Sibutramine (10mg/kg/day p.o., 4 weeks respectively) was administered followed by
administration of sulpiride (20 mg/kg i.p.) after a gap of 2 h for 28 consecutive days.
Group X: Ethanolic extract of Vitis vinifera (250mg/kg p.o. respectively) was administered
followed by administration of sulpiride (20 mg/kg i.p.) after a gap of 2 h for 28 consecutive days.
Group XI: Ethanolic extract of Vitis vinifera (500mg/kg p.o. respectively) was administered
followed by administration of sulpiride (20 mg/kg i.p.) after a gap of 2 h for 28 consecutive days.
Group XII: Ethanolic extract of Solanum lycopersicum (250mg/kg p.o. respectively) was
administered followed by administration of sulpiride (20 mg/kg i.p.) after a gap of 2 h for 28
consecutive days.
Group XIII: Ethanolic extract of Solanum lycopersicum (500mg/kg p.o. respectively) was
administered followed by administration of sulpiride (20 mg/kg i.p.) after a gap of 2 h for 28
consecutive days.
Results
Parameters
Standard diet
High fat diet
Sulpride
Serum Cholesterol (mg/dl)
52.86 ± 3.03
126.77 ± 7.41a
84.31± 7.96b
Serum Triglyceride (mg/dl)
58.46 ± 4.90
160.73 ± 9.29a
103.94 ± 5.86b
Serum HDL (mg/dl)
31.28 ± 3.14
15.74 ± 4.52a
19.84 ± 4.52b
Serum LDL (mg/dl)
7.87 ± 0.51
20.44 ± 1.12a
19.84 ± 1.52b
Table 4.3- Effect of standard diet, high fat diet and Sulpride on serum lipid profile. ALL Values
are represented as mean ± S.D; a = P < 0.05 vs. standard diet Control on Day 56 and b= P < 0.05
vs. standard diet Control on Day 28th day.
Parameters
Standard diet
High fat diet
Sulpride
TBARS (nmol/mg protein)
0.582± 0.068
1.385 ± 0.135a
1.071±0.120b
Reduced glutathione (nmol/mg 21.55 ± 1.51
9.45 ± 0.72a
10.71 ± 1.52b
protein)
Catalase (µmol of H2O2 0.709 ± 0.041
0.147 ± 0.022a
0.167 ± 0.032b
decomposed/min/mg
of
protein)
Super
oxide
dismutase 52.86 ± 3.03
13.87 ± 1.11a
17.32 ± 1.56b
units/mg of protein
Table 4.4- Effect of standard diet, high fat diet and Sulpride on TBARS and different antioxidant
enzymes ALL Values are represented as mean ± S.D; a = P < 0.05 vs. standard diet Control on
Day 56 and b= P < 0.05 vs. standard diet Control on Day 28th day
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Fig- 4.1 Effect of various pharmacological interventions on serum total cholesterol. ALL Values
are represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P < 0.05 vs. HFD on 56
Day., c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat diet., SIBSibutramine., SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis vinifera
(500mg/kg/day)., TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum lycopersicum
(500mg/kg/day).
Fig- 4.2 Effect of various pharmacological interventions on serum triglyceride. ALL Values are
represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P < 0.05 vs. HFD on 56 Day.,
c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat diet., SIB- Sibutramine.,
SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis vinifera (500mg/kg/day).,
TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum lycopersicum (500mg/kg/day).
S eru m H D L (m g/d l)
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Fig- 4.3 Effect of various pharmacological interventions on serum high density lipoprotein. ALL
Values are represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P < 0.05 vs. HFD
on 56 Day., c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat diet., SIBSibutramine., SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis vinifera
(500mg/kg/day)., TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum lycopersicum
(500mg/kg/day).
25
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Fig- 4.4 Effect of various pharmacological interventions on serum low density lipoprotein. ALL
Values are represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P < 0.05 vs. HFD
on 56 Day., c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat diet., SIBSibutramine., SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis vinifera
(500mg/kg/day)., TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum lycopersicum
(500mg/kg/day).
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R educed glutathione (nm ol/m g protein).
Fig- 4.5 Effect of various pharmacological interventions on lipid peroxidation and antioxidant
enzmes. ALL Values are represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P <
0.05 vs. HFD on 56 Day., c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat
diet., SIB- Sibutramine., SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis
vinifera (500mg/kg/day)., TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum
lycopersicum (500mg/kg/day).
Fig- 4.6 Effect of various pharmacological interventions on enzyme reduced glutathione. ALL
Values are represented as mean ± S.D; a = P < 0.05 vs. Normal Control., b = P < 0.05 vs. HFD
on 56 Day., c =P < 0.05 vs. Sulpride Control treatment on 28 Day; HFD- high fat diet., SIBSibutramine., SUL- Sulpride., GLD- Vitis vinifera (250mg/kg/day)., GHD- Vitis vinifera
(500mg/kg/day)., TLD- Solanum lycopersicum(250mg/kg/day)., THD-Solanum lycopersicum
(500mg/kg/day).
Disscusion
Obesity is caused by low energy expenditure and increased fatty acid synthesis from
carbohydrates and fat intake by organs. In hamsters, tomato lowered the cholesterol diet-induced
levels of plasma cholesterol, LDL-cholesterol and triglyceride [13]. Unlike red tomato, green
tomato contains high amounts of α-tomatine, dehydrotomatine and trigonelline and low amounts
of carotenoids [14]. A tomatine enriched diet significantly decreased plasma LDL-cholesterol
levels in hamsters fed a high-fat, high-cholesterol diet [14]. Tomatine also reduced serum levels
of cholesterol and triglyceride and liver cholesterol levels in cholesterol-fed rats [15]. Grape seed
extract lowered the cholesterol diet-induced levels of plasma cholesterol, LDL-cholesterol and
triglyceride Grape ingredients also have healthful properties.
Sibutramine treatment significantly decreases (p<0.05) in total cholesterol, serum
triglyceride, serum low density lipoprotein and increases in high density lipoprotein in mg/dl as
compared with high fat diet control in 8 weeks and sulpride control in 4 weeks (Table no- 4.9
Fig no- 4.13, 4.14, 4.15, 4.16). Treatment with Vitis vinifera and Solanum lycopersicum
produced significant decrease (p<0.05) in total cholesterol, serum triglyceride, serum low
density lipoprotein and increases in high density lipoprotein as compared to HFD group in 8
weeks Sulpride control group in 4 weeks (Table no- 4.9 Fig no- 4.13, 4.14, 4.15, 4.16). High fat
diet and sulpiride significantly decreases the antioxidant enzymes and increases the lipid
peroxidation as the result shown in (Table No-.4.6). Antioxidant enzymes reverse the lipid
profile of Obesity [16] (Thamolwan et al., 2010). So that findings clarify that various dosage of
ethanolic extract of Vitis vinifera and Solanum lycopersicum suppresses the obesity caused by
the High fat diet and Sulpiride.
High fat diet significantly increases the tissue TBARS levels and decreases antioxidant
enzymes i.e. reduced glutathione, catalase, super oxide dismutase in the rats on high fat diet for 8
weeks [17] on the other hand sulpiride significantly increases the levels of TBARS and decreases
antioxidant enzymes i.e. reduced glutathione, catalase, super oxide dismutase in the liver tissue
in 4 weeks [18].
A significant Increase (p<0.05) in tissue concentration of lipid peroxidation i.e TBARS
and significant (p<0.05) decrease in tissue antioxidant enzymes i.e. catalase, reduced
glutathione, and super oxide dismutase concentration was observed in rats after 8 week of high
fat diet and 4 weeks of sulpride (Table no- 4.6, Fig. No.4.17, 4.20). Sibutramine treatment
significantly decreases (p<0.05) tissue concentration of lipid peroxidation i.e. TBARS in
nmol/mg of protein and increases in tissue antioxidant enzymes i.e. catalase, reduced
glutathione, and super oxide dismutase concentration as compared with high fat diet control in
8 weeks sulpride control in 4 weeks (Table no- 4.10, Fig no- 4.18, 4.19, 4.21, 4.22). Treatment
with ethanolic extract of various doses of Vitis vinifera and Solanum lycopersicum produced
significant decrease (p<0.05) in lipid peroxidation i.e. TBARS in nmol/mg of protein and
increases in tissue antioxidant enzymes i.e. catalase, reduced glutathione, and super oxide
dismutase concentration as compared to HFD group in 8 weeks and Sulpride control group in 4
weeks (Table no- 4.10, Fig no- 4.18, 4.19, 4.21, 4.22). These findings clarify that various dosage
of ethanolic extract of Vitis vinifera and Solanum lycopersicum suppresses the obesity caused by
the High fat diet and Sulpiride.
Summary and Conclusion
The salient findings of the present study may be summarized as follows:
1. High fat diet and sulpiride has produced significantly increase serum lipid (total cholesterol,
LDL, VLDL, and triglycerides) and decreases the serum HDL to produce experimental
obesity.
2. Vitis vinifera (250mg/kg/day), Vitis vinifera (500mg/kg/day), Solanum lycopersicum
(250mg/kg/day), Solanum lycopersicum (500mg/kg/day) significantly decreases serum lipid
(total cholesterol, LDL, VLDL, and triglycerides) and increases the serum HDL to produce
antiobesity effect.
3. High fat diet and sulpiride has produced significantly increase in tissue parameters i.e.
TBARS and significantly decreased the tissue SOD, Catalase, Super oxide dismutase to
produce experimental obesity.
4. Vitis vinifera (250mg/kg/day), Vitis vinifera (500mg/kg/day), Solanum lycopersicum
(250mg/kg/day), Solanum lycopersicum (500mg/kg/day) significantly decreases the tissue
TBARS and significantly increases the tissue SOD, Catalase, Super oxide dismutase to
produce antiobesity effect.
5. The antiobesity effect of various doses of ethanolic extract of Vitis vinifera and Solanum
lycopersicum is produced due to the antioxidant effect produced from the both plants.
Reference
1. Eikelis N, and Elser M: The neurobiology of human obesity. Exp. Physiol 2005;
90:5673–682.
2. Chaput JP, and Tremblay :A Current and novel approaches to the drug therapy of obesity;
Eur J Clin Pharmacology2006; 62(10):793-803.
3. Yoo MA, Chung HK and Kang MH :Evaluation of physicochemical properties in
different cultivar grape seed waste; Food Sci Biotechnol 2004; 13:26-29.
4. Shi J, Yu J, Pohorly JE and Kakuda Y :Polyphenolics in grape seeds-biochemistry and
functionality; J Med Foo 2003; 6:291-299.
5. Jang JK & Han JY :The antioxidant ability of grape seed extracts; Korean Journal of
Food Science and Technology 2002; 34:524-528.
6. Srinivasan K, Viswanad B, Asrat L, Kaul CL, and Ramarao P :Combination of high-fat
diet-fed and low-dose streptozotocin-treated rat: A model for type 2 diabetes and
pharmacological screening; Pharmacol Res 200; 52: 313–320.
7. Baptista T, Q Contreras, L Teneud, M Quijada, A Acosta, and A LaCruz :Mechanism of
the neuroleptic-induced obesity in female rats; Prog. Neuropsychopharmacol. & Biol.
Psychiat 1997; 22: 187-198.
8. Allain CC, Poon LS, Chan CS, Richmond W and Fu .C :Enzymatic determination of total
serum cholesterol; Clin. Chem 1974; 20: 470-475.
9. Werner M, Gabrielson DG and Eastman J :Ultramicro determination of serum
triglycerides by bioluminescent assay; Clin. Chem 1981; 27: 268-271.
10. Friedewald WT, Levy RI, and Fredrickson DS :Estimation of the concentration of lowdensity lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge;
Clin. Chem 1972; 18: 499-502.
11. Ohkawa H, Ohishi N, Yagi K :Assay for lipid peroxides in animal tissue by thiobarbituric
acid reaction; Anal Biochem 1979; 95: 351-358.
12. Ellman, GL :Tissue sulfhydryl groups; Arch Biochem Biophys 1959; 82: 48670-48677.
13. Friedman M :Tomato glycoalkaloids: role in the plant and in the diet; J Agric Food Chem
2002 ;50: 5751-80.
14. Leonardi C, Ambrosino P, Esposito F, Fogliano V :Antioxidative activity and carotenoid
and tomatine contents in different typologies of fresh consumption tomatoes; J Agric
Food Chem 2000; 48:4723-7.
15. Cayen MN :Effect of dietary tomatine on cholesterol metabolism in the rat; J Lipid Res
1971; 12:482-90.
16. Thamolwan S, Watcharaporn D, Thanapat S, Suwan T, Somlak P :Antioxidant Activity
and Lipid-Lowering Effect of Essential Oils Extracted from Ocimum sanctum L. Leaves
in Rats Fed with a High Cholesterol Diet; J Clin Biochem Nutr 2010; 46(1): 52–59.
17. Mohammed AH,:Anti-obesity, antiatherogenic, anti-diabetic and antioxidant activities of
J. montana ethanolic formulation in obese diabetic rats fed high-fat diet; Free RAD.
Antiox 2011; 1: 49-60.
18. Dietrich MA, Olas B :Inhibitory effects of polyphenol compounds on lipid peroxidation
caused by antipsychotics (haloperidol and amisulpride) in human plasma in vitro; World
journel of psychiatry 2010; 2:276-81.