Fitoterapia 77 (2006) 160 163 www.elsevier.

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Beneficial effects of Zingiber officinale on goldthioglucose induced obesity

Ramesh K. Goyal , Sanjay V. Kadnur
Department of Pharmacology, L.M.College of Pharmacy, Navarangpura, Ahmedabad-380009, India Received 14 February 2005; accepted 10 January 2006 Available online 28 February 2006

Abstract Goldthioglucose induces in mice a significant increase in body weight, glucose, insulin and lipid levels. Treatment with 250mg/ kg of methanol and ethyl acetate extracts of Zingiber officinale for 8 weeks produces significant reduction in body weight, glucose, insulin and lipid levels as compared to obese control mice. The reduction in elevated glucose along with elevated insulin levels indicates that the treatment with Z. officinale improves insulin sensitivity. 2006 Published by Elsevier B.V.
Keywords: Zingiber officinale; Obesity; Goldthioglucose

1. Introduction The prevalence of obesity in the United States and world wide is increasing. More than one half of U.S. men and women aged 20 years are considered over weight [Body mass Index (BMI) in kg/m2 25] and nearly one fourth are clinically obese [BMI in kg/m2 30]. A moderate obesity can contribute to chronic metabolic abnormalities characteristic of insulin resistance syndrome, such as dyslipidemia, hypertension, insulin resistance and glucose intolerance [1]. Environmental factors such as the general availability of high calorie food or the limited need for physical exercise and genetic factors that predispose to weight gain contribute to the development of obesity [2]. Drugs like fenfluramine and dexfenfluramine used in earlier days for treating obesity act by modulating 5-hydroxytryptamine (5-HT) actions like release of 5-HT, inhibition of its reuptake or both. Sibutramine an inhibitor of both norepinephrine reuptake and serotonin that also weakly inhibits dopamine reuptake is used in the treatment of obesity [3]. 5-HT plays a crucial role in controlling appetite, hence drugs modulating 5-HT functions are among good candidates for screening for their efficacy in obesity. Zingiber officinale commonly known as ginger is one of the commonly used spices in India and around the world. Ginger is reported to have various pharmacological activities like antiemetic, antiulcer, anxiolytic, antiinflammatory and antipyretic activities [49]. Majority of the pharmacological activities reported for Z. officinale can be explained

Corresponding author. Tel.: +91 79 26302746; fax: +91 79 26304865. E-mail address: goyalrk@hotmail.com (R.K. Goyal). 0367-326X/$ - see front matter 2006 Published by Elsevier B.V. doi:10.1016/j.fitote.2006.01.005

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by its effect on 5-HT receptor modulation. Earlier studies from our laboratory had demonstrated that fresh juice of Z. officinale reduces the elevated lipid levels associated with hyperglycemia in streptozotocin-induced type I diabetic in rats. It was also correlated that reduction in the serum glucose levels are mediated by 5-HT receptor antagonism [10]. However, in our earlier studies reduction in serum glucose levels were seen in conditions of hypoinsulinemia. In the present study Z. officinale is evaluated for its beneficial effects in conditions of hyperglycemia associated with hyperinsulinemia, hyperlipidemia and increased abdominal fat deposition which represents obesity. 2. Experimental 2.1. Plant material Dried rhizomes of Z. officinale Roscoe (Zingiberaceae) were purchased from a local market. They were identified by morphologic and microscopic comparison according to different standard texts by Prof. O. P. Saxena, Head of the Botany Department, Gujarat University, Ahmedabad, India. A specimen was deposited in the Botany Department, Gujarat University, Ahmedabad, India. 2.2. Preparation of methanolic and ethyl acetate extracts Dried and powder rhizomes were extracted with MeOH under reflux for 6h. After filtration and concentration in vacuo the methanolic extract was obtained (yield: 4.3%). The AcOEt extract was obtained in the same condition (yield: 4.6%). 2.3. Animals Female Swiss mice of 4 weeks were used. They were housed in a standard environmental conditions and fed with standard rodent diet and water ad libitum. They were initially acclimatized for the study and the study protocol was approved by the Institutional Animal Ethics Committee of our college as per the requirements of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), New Delhi, India. 2.4. Experimental induction of obesity in mice Animals were administered goldthioglucose (Sigma, USA) at a dose of 300mg/kg i.p. Animals after attaining 12 18 weeks of age were checked for development of obesity which was characterized by increased body weight (animals weighing above 30 g), abdominal fat deposition and higher lipid profile when compared to control mice. Age matched control mice which received normal saline at 4weeks of age on attaining 1218 weeks were selected for the study as normal control [11]. A total of 24 mice were equally divided into 4 groups (n = 6 in each group). Group I: normal control (received vehicle). Group II: obese mice (received vehicle). Group III: obese mice treated with methanolic extract of Z. officinale (250 mg/kg p.o.). Group IV: obese mice treated with ethyl acetate extract of Z. officinale (250 mg/kg p.o.). The extracts were administered for 8 weeks. Daily, body weight of animals were recorded. At the end of the treatment period, animals were kept for overnight fasting. Blood samples were then collected from the tail vein. The
Table 1 Effects of Z. officinale methanolic and ethyl acetate extracts on body weight, serum glucose and insulin levels on goldthioglucose-induced obesity in mice Dose Normal control (vehicle) Obesity control (OB) (vehicle) OB + methanolic extract (250mg/kg) OB + ethyl acetate extract (250mg/kg) N= 6. Values are mean S.E.M. a Significantly different from normal control P b 0.05. b Significantly different from obesity control P b 0.05. Body (g/mice) 22.5 1.1 41.6 1.6 a 32.5 1.1 b 35.8 0.8 b Glucose (mg/dl) 78.5 1.1 156.7 1.7 a 124.4 2.5 b 128.7 2.1 b Insulin U/ml 31.0 0.8 91.3 2.1 a 75.0 1.5 b 88.3 1.8 b

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Table 2 Effects of Z. officinale methanolic and ethyl acetate extracts on serum lipid profile on goldthioglucose-induced obesity in mice Dose Normal control OB control OB + methanolic extract (250 mg/kg) OB + ethyl acetate extract (250 mg/kg) Cholesterol (mg/dl) 61.9 0.9 107.9 1.8 a 67.4 2.4 b 66.8 1.6 b Triglycerides (mg/dl) 35.6 0.8 78.3 0.6 a 36.0 1.0 b 40.9 0.5 b HDL-cholesterol (mg/dl) 11.2 0.3 12.0 0.5 11.1 0.6 11.2 0.4 LDL-cholesterol (mg/dl) 43.3 0.9 80.1 2.3 a 49.0 2.1 b 47.4 1.7 b VLDL-cholesterol (mg/dl) 7.1 0.1 15.6 0.1 a 7.2 0.2 b 8.1 0.1 b

N = 6.Values are mean S.E.M. a Significantly different from normal control P b 0.05. b Significantly different from obesity control P b 0.05.

blood collected was allowed to clot for 30min at r.t., blood samples were centrifuged at 5000 rev./min for 15 min. Serum samples thus obtained were stored at 20 C until biochemical estimations were carried out. 2.5. Biochemical analysis The serum samples, obtained as mentioned above, were analyzed for the following biochemical parameters: serum glucose by GODPOD method, cholesterol by cholesterol esterase method, triglyceride by Glycerol-3-Phosphate Oxidase method, HDL-cholesterol by phosphotungstate method using respective diagnostic kits obtained from Bayer Diagnostics Ltd, India. Serum insulin levels were estimated by radioimmunoassay method using the kit from Bhabha Atomic Research Center, Mumbai, India. VLDL-Cholesterol and LDL-Cholesterol were calculated as per Friedevald's equation. VLDL Cholesterol Total serum triglycerides 5 Total serum triglycerides HDL C LDL C Total serum cholesterol 5

2.6. Statistical analysis Result were analyzed statistically using analysis of variance (ANOVA) followed by Tukey's test. Values of P b 0.05 were considered significant. 3. Results Animals of group II (obese control) showed significant increase in body weight, serum glucose and insulin levels as compared to normal control mice. Treatment with methanolic and ethyl acetate extracts significantly reduced body weight from 41.6 to 32.5 and 35.8 g, respectively. The treatment significantly reduced the serum glucose and insulin levels (Table 1). Obese control mice (group II) also showed significant increase in serum cholesterol, triglyceride, LDL-cholesterol, VLDL-cholesterol levels and no change in serum HDL-cholesterol levels as compared to normal mice. Treatment with methanolic and ethyl acetate extracts produced significant reduction in serum cholesterol, triglyceride, LDLcholesterol, VLDL-cholesterol. No change in serum HDL-cholesterol levels was observed as compared to obese control mice (Table 2). 4. Discussion Goldthioglucose induced obesity model is one among the accepted models for screening of drugs useful in obesity. Goldthioglucose induces destruction of hypothalamic and extra hypothalamic areas of the brain and induces obesity [1114]. In the present study, obese control mice showed significantly higher levels of fasting glucose and insulin levels as compared to control mice. This is consistent with earlier reports [1114]. Treatment with methanolic and ethyl acetate extracts of Z. officinale to obese mice, produced a significant reduction in serum glucose levels. We have previously shown these findings with fresh juice of ginger in diabetic animals [10]. Treatment with Z. officinale methanolic and

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ethyl acetate extracts to obese mice produced a significant reduction in insulin levels and glucose levels. This suggests that the antihyperglycemic effect produced is an extra pancreatic effect, enhancing the insulin sensitivity. Treatment with extracts of Z. officinale shows reduction in elevated lipid levels associated with hyperglycemia and hyperinsulinemia. The reduction in elevated lipid levels is supported by earlier reports which have shown that treatment with Z. officinale in hypercholesteremic animal models, produces a significant reduction in elevated lipid levels [5,6]. As reported earlier, insulin resistance or insulin deficiency is associated with hypercholesterolemia and hypertriglyceredemia. Insulin resistance may be responsible for dyslipidemia, because insulin has an inhibitory action on HMG-CoA reductase, a key rate limiting enzyme responsible for the metabolism of cholesterol rich LDL particles [15]. The reduction in glucose and elevated lipid levels in the condition of obesity associated with insulin resistance involving hyperglycemia and hyperinsulinemia indicates that Z. officinale improves insulin sensitivity. 5-HT receptor modulators are reported to have beneficial effects in conditions of obesity. 5-HT2C receptors which are subtype of 5-HT receptors is restricted to CNS and this receptor is responsible for anorectic properties of 5-HT [2]. Sibutramine an inhibitor of both norepinephrine reuptake and serotonin that also weakly inhibits dopamine reuptake is used in the treatment of obesity [3]. 5-HT plays a crucial role in controlling appetite, hence drugs modulating 5-HT functions and thereby control the appetite are useful in conditions of obesity. Z. officinale is reported to contain 5-HT modulators [16]. The antianxiety and antiemetic effects of Z. officinale are mediated by 5-HT receptors [7]. Earlier studies from our laboratory reported antidiabetic activity with fresh juice of ginger in diabetic animals. Our earlier study also showed the antidiabetic activity of ginger mediated through 5-HT receptor antagonism [10]. Observations found in this study seems to provide enough support to suggest the effects of Z. officinale in reducing elevated glycemic levels, lipid levels and reduction in body weight as compared to obese control mice may be due to its 5-HT modulatory effects. In conclusion our data suggest that methanolic and ethyl acetate extracts of Z. officinale has potential beneficial effects in conditions of obesity. In addition to decrease serum glucose, insulin and lipid levels it can also significantly retard gain in body weight. Based on these beneficial effects, Z. officinale can be considered as supplementary herbal therapy in obese patients for prevention or treatment of obesity. Acknowledgements Authors would like to acknowledge University Grants Commission, New Delhi for financial aid under Major Research Project No. F-7-40/2001(SR-I) to Dr. Goyal with Mr. Kadnur as the Project Associate. References
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