Journal of Ethnopharmacology 88 (2003) 107111

Antihyperglycemic effects of three extracts from Momordica charantia

Jaspreet Virdi a , S. Sivakami a , S. Shahani b , A.C. Suthar c , M.M. Banavalikar c, , M.K. Biyani c
c

Department of Life Science, University of Mumbai, Kalina, Santacruz (East), Mumbai 400 098, India b Department of Pharmacology, LTMMG Hospital, Sion, Mumbai 400 022, India Ajanta Pharma Ltd., Research Centre, Ajanta House, 98, Govt. Industrial Estate, Charkop, Kandivali (West), Mumbai 400 067, India Received 17 May 2002; received in revised form 13 May 2003; accepted 21 May 2003

Abstract Momordica charantia (L.) (Cucurbitaceae) commonly known as bitter gourd or karela is a medicinal plant, used in Ayurveda for treating various diseases, one of which is diabetes mellitus. In this study, various extract powders of the fresh and dried whole fruits were prepared and their blood glucose lowering effect compared by administrating them orally to diabetic rats. The aqueous extract powder of fresh unripe whole fruits at a dose of 20 mg/kg body weight was found to reduce fasting blood glucose by 48%, an effect comparable to that of glibenclamide, a known synthetic drug. This extract was tested for nephrotoxicity, hepatotoxicity and biochemical parameters such as SGOT, SGPT and lipid prole. The extract did not show any signs of nephrotoxicity and hepatotoxicity as judged by histological and biochemical parameters. Thus the aqueous extract powder of Momordica charantia, an edible vegetable, appears to be a safe alternative to reducing blood glucose. 2003 Elsevier Ireland Ltd. All rights reserved.
Keywords: Momordica charantia; Karela; Bitter gourd; Hypoglycemic activity; Nephrotoxicity; Hepatotoxicity

1. Introduction Diabetes is a disorder of carbohydrate, fat and protein metabolism attributed to diminished production of insulin or mounting resistance to its action. Chronic hyperglycemia during diabetes causes glycation of body proteins that in turn leads to secondary complications affecting eyes, kidneys, nerves and arteries (Sharma et al., 1993). These may be delayed, lessened or prevented by maintaining blood glucose values close to normal. Besides the use of insulin for the treatment of insulin dependent diabetes mellitus (IDDM), other approaches for the control of hyperglycemia include the use of amylin analogues which regulate gastric emptying and inhibitors of intestinal alpha glucosidases like acarbose, miglitol and voglibiose which delay postprandial hyperglycemia. Sulphonylureas, the most widely used class of drugs act by closure of ATP dependent channel. Metformin, a biguanide oral antibiotic limits intestinal glucose absorption. These drugs have certain effects like causing hypoglycemia at higher doses, liver problems, lactic acidosis and diarrhea. It is apparent that due to the side effects of the cur Corresponding author. Tel.: +91-86-83945/625/718; fax: +91-86-83930/2845. E-mail addresses: manishab@ajantapharma.com, mbanavalikar@hotmail.com (M.M. Banavalikar).

rently used drugs, there is a need for a safe agent with minimal adverse effects, which can be taken for long durations. In addition to the above drugs of synthetic origin, many agents of plant origin are also in use particularly for the treatment of non-insulin dependent diabetes mellitus (NIDDM). Various medicinal plants like Momordica charantia L., Azardiracta indica and Ficus racemosa are known to possess antihyperglycemic activity (Atta-ur-Rahman, 1989). Extracts of the leaves of Gymnema sylvestris have been shown to induce beta cell regeneration (Baskaran et al., 1990). A galactomannan has been identied as the major constituent of the blood glucose lowering extract from Trigonella foenum-graecum (Ali et al., 1995). The hypoglycemia producing molecule of Pandanus odorus roots has been shown to be 4-hydroxy benzoic acid (Peungvicha et al., 1998). A pectin from the fruit of Coccinia indica signicantly lowered blood glucose (Kumar et al., 1993). For the present study Momordica charantia (L.) (Cucurbitaceae) was chosen since it is by far the most extensively investigated and most widely acclaimed remedy for treatment of diabetes mellitus since ancient times. Momordica charantia, also referred to as bitter gourd or karela, is a member of the Cucurbitaceae family and is commonly used as a traditional remedy for diabetes in India, Asia, Africa and South America. It is commonly consumed as a vegetable in India. The fruit, leaves, seeds and roots of Momordica

0378-8741/$ see front matter 2003 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/S0378-8741(03)00184-3

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charantia have been used in the Indian system of medicine for a number of diseases, besides diabetes. The unripe fruits of this plant have been shown to produce a hypoglycemic effect in experimental models following oral administration (Sharma et al., 1960; Akhtar et al., 1981; Srivastava et al., 1987; Day et al., 1990; Pugazhenthi et al., 1995). Many clinical trials have also conrmed the hypoglycemic action (Leatherdale et al., 1981). Some investigators have attempted to purify the active fractions from fruits of Momordica charantia in order to establish the precise mechanism of its hypoglycemic effects (Lotlikar and Rajarama Rao, 1966). But, most of these studies were restricted to very short durations and used a single large dose. In the present study, extracts of the fresh and dried whole fruits were compared for their efcacies in lowering fasting blood glucose. The extract, which showed the maximum efcacy, was further investigated for hepato and nephrotoxicity during the experimental period.

glucose levels above 250 mg%, were considered diabetic and selected for the study. The urine glucose was monitored using glucotest strips. 2.3. Preparation of the extracts 2.3.1. Extract A This was prepared by extracting 0.5 kg of dried whole fruits of Momordica charantia along with the seeds using methanol in a ratio of 1:10. The extraction was carried out at 50 C for 1 h with stirring at regular intervals. It was then ltered and evaporated to dryness under reduced pressure to get 5.6% bitter content. The yield was 37 g. 2.3.2. Extract B This was prepared by extracting 0.5 kg of dried whole fruits of Momordica charantia along with seeds using chloroform in ratio of 1:10. The extraction was carried out exactly as described above. The extract had 4.8% bitter content. The yield was 28 g. 2.3.3. Extract C This was prepared by extracting 0.5 kg of fresh green whole fruits of Momordica charantia along with seeds using water at a ratio of 10:25. Pieces of fruits were soaked in water for 1 h at room temperature. It was then ltered and evaporated to dryness under reduced pressure to get 4.1% bitter content. The yield was 10 g. 2.4. Mode of feeding The rats were administered the materials twice a day for a period of 4 weeks. The extract powders were orally fed at a dosage of 20 mg/kg body weight and glibenclamide at a dosage of 0.1 mg/kg body weight. The rats were weighed everyday throughout the study. Fasting blood was collected from the tail vein once a week and glucose estimated. In the case of glibenclamide and that extract which shows the best hyperglycemic activity, blood was collected from the animals by retro-orbital bleeding, at the end of the 4th week, sera separated and used for the determination of biochemical parameters, cholesterol, HDL, VLDL, triglycerides, SGOT, SGPT, creatinine and uric acid. 2.5. Fixation and staining of tissues

2. Methods and materials Green, unriped fresh fruits of Momordica charantia were purchased from the local market of Mumbai, India and identied by Blatter Herbarium, St. Xavier College, Mumbai, India. A voucher specimen (No. 5475 of Northern Island) has been kept in our laboratory for future reference. Part of fresh fruits were sun dried to get Momordica charantia fruit dried powder. Alloxan and glibenclamide were obtained from Sigma Co., St. Louis, MO, USA. The glucose OxidasePeroxidase kits was purchased from Span Diagnostics, India. The glucotest urine strips were obtained from Bayer Diagnostics India Ltd. 2.1. Experimental animals Males Wistar rats weighing 150180 g were purchased from Haffkine BioPharma Ltd., Mumbai. The rats were housed in air-conditioned animal house. Each rat was kept in a separate cage and fed Amrut rat chow and boiled water ad libitum. 2.2. Induction of diabetes After acclimatization, the rats were weighed and divided into the following groups of six animals each: untreated controls, untreated diabetics, controls fed with extracts, diabetics fed with extracts. The control group was injected 0.85% saline intraperitoneally. Diabetes was induced by intraperitoneal injection of alloxan at a dose of 175 mg/kg body weight. The alloxan solution was freshly prepared, kept on ice and injected immediately. Non-fasting blood samples were collected via tail vein and used for measuring blood glucose levels by the Glucose OxidasePeroxidase (GOD/POD) method. Only those rats, which showed blood

At the end of 4 weeks the rats were sacriced and livers and kidneys collected to detect hepatotoxicity and nephrotoxicity. The tissues were xed in 10% formalin immediately after removal from the animal to avoid decomposition. Embedding in parafn wax was carried out by removal of water using alcohol dehydration and inltration of xylene or chloroform as a solvent for the wax. Thin sections of the tissue (7 m) were cut using a microtome after the solidication of parafn wax and stained with hematoxylin eosin. The tissue sections were subjected

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to rehydration by exposure to decreasing concentrations of alcohol from 100 to 30% and then stained with hematoxylin, which has an aqueous base. The sections were dehydrated using increasing concentrations of alcohol followed by staining with eosin. They were then treated with diphenyl pthalate xylene (DPX) and observed under the microscope (Pearse, 1981). 2.6. Statistical analysis Data are expressed as mean S.D.

3. Results and discussion As can seen from Table 1, though all three extract powders did lower blood glucose, the aqueous extract powder showed the maximum efcacy. The methanolic extract (A) reduced blood glucose by 49% at the end of the rst week which became 39% at the end of the fourth week. While the chloroform extract (B) showed almost no antihyperglycemic activity (3%), the aqueous extract (C) showed a blood glucose lowering activity, which was 50% and stayed consistent till the end of the study. The antihyperglycemic activity of extract C was comparable to that of glibenclamide. The extracts A and C also lowered blood glucose by 24 and 18% in controls, respectively (Table 1). The superior effects of the water extract from the fresh fruit is in keeping with the observation of Leatherdale et al. (1981) that dried bitter gourd was less effective than aqueous extracts in lowering blood glucose. Methanol was used for extract A keeping in mind that certain active principles like phytosterols, can be extracted with methanol. However, the methanolic extract from the dried fruits was less effective in controlling blood glucose. These observations also gain signicance due to the fact that whole bitter gourd is cooked in water and consumed in many cultures, particularly in India. The oral feeding of extract A for 4 weeks resulted in an increase in body weight which was 6 and 23% in diabetic

and control animals, respectively. The extract B also led to a gain in body weight being 4% in diabetics and 25% in controls. An increase in body weight of 4% in diabetics and 21% in controls was seen upon administration of the extract C. In the case of glibenclamide, there was no net gain in the weight of diabetic animals, but the controls showed a 24% increase. Thus the diabetic animals fed the extracts showed less weight gain than the corresponding controls. This observation and the decrease in body weight observed in uncontrolled diabetics might be the result of protein wasting due to unavailability of carbohydrate for utilization as an energy source (Table 2). Control animals fed with the extracts A, B, C and glibenclamide do not excrete any glucose in urine excluding any harmful effect of the extracts on the kidneys. But, none of these extracts were able to inhibit or reverse to any significant extent the excretion of glucose in the urine of the diabetics. Considering the above results obtained, extract C was chosen for a further detailed study of biochemical parameters. Glibenclamide, a synthetic drug, was used as a reference for comparison. Abnormalities in lipoproteins are very common in both NIDDM and IDDM. Although lipoprotein alterations appear to be an intrinsic part of these disorders, such alterations are also induced by diabetes associated complications such as obesity and renal disease. In the present study, total cholesterol along with HDL levels remained unchanged in diabetics compared to the other groups. The triglyceride levels were observed to be elevated in untreated diabetics but reduced by both extract C (69% in diabetics and 32% in controls) as well as glibenclamide (65% in diabetics and 18% in controls) showing their benecial effects. The elevated VLDL levels in the untreated diabetics were also reduced by extract C and glibenclamide. These results suggest the benecial effects of the natural extract in improving the imbalance in lipoprotein metabolism are also comparable to those of glibenclamide. Kidneys maintain optimum chemical composition of body uids by acidication of urine and removal of metabolite

Table 1 Comparison of efcacy of different extracts of Momordica charantia with glibenclamide for percentage decrease in blood glucose Group Duration of treatment (in weeks) 1st week Diabetic + extract A Control + extract A Diabetic + extract B Control + extract B Diabetic + extract C Control + extract C Diabetic + glibenclamide Control + glibenclamide 49.2 24.2 3.9 10.0 49.0 21.8 50.8 23.5 1.4 6.0 1.9 0.6 2.7 2.4 0.5 2.6 2nd week 47.0 17.6 3.8 6.9 50.0 21.4 52.0 20.8 1.3 2.2 0.1 0.4 1.5 4.3 1.3 5.9 3rd week 46.1 20.3 3.7 9.8 51.7 21.1 51.1 21.7 1.4 1.5 1.6 0.7 1.7 3.3 0.5 2.4 4th week 39.0 26.8 3.8 13.9 50.8 18.2 51.5 18.2 3.7 1.2 1.8 4.5 0.9 0.8 1.5 0.6

Blood glucose values (mg%) of untreated controls and diabetic Diabetic 270.9 1.9 Untreated control 110.0 0.6

295.8 2.7 106.9 0.4

324.7 1.6 109.8 0.7

359.8 1.8 113.9 4.5

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Table 2 Variation in body weights of animals treated with different extracts of Momordica charantia and glibenclamide (in grams) Group Duration of treatment (in weeks) 1st week Diabetic + extract A Control + extract A Diabetic + extract B Control + extract B Diabetic + extract C Control + extract C Diabetic + glibenclamide Control + glibenclamide 201.1 239.2 185.2 215.8 187.5 227.0 197.7 225.0 7.4 4.7 1.6 1.1 7.3 0.6 10.3 6.2 2nd week 189.1 220.8 170.0 220.4 194.0 241.7 203.6 252.1 7.9 3.6 4.6 2.9 8.3 12.8 8.5 7.7 3rd week 193.6 263.8 171.6 238.8 198.6 272.9 203.4 272.5 7.5 7.7 4.3 5.9 4.1 4.1 6.2 9.4 4th week 215.8 295.8 193.9 270.0 194.2 275.8 198.8 281.2 8.0 1.1 4.2 5.2 9.1 4.7 6.3 7.6

Mean weight (in grams) of untreated control and diabetic Diabetic 215.9 7.9 Untreated control 221.6 4.5

208.8 12.7 252.9 9.3

203.7 11.6 269.8 7.0

202.8 9.8 272.9 8.5

Table 3 Determination of biochemical parameters after treatment with Momordica charantia extract C and glibenclamide Diabetic + extract C Control + extract C Diabetic + glibenclamide Control + glibenclamide Diabetic Total cholesterol (mg/dl) 67.37 1.5 HDL cholesterol (mg/dl) 57.6 2.2 Triglycerides (mg/dl) 134.3 1.8 VLDL 27.8 1.4 SGOT (U/l) 252.5 1.8 SGPT (U/l) 97.5 1.3 Creatinine 4.8 0.3 Uric acid 3.4 0.2 52.27 2.1 49.5 1.0 133.3 4.2 27.6 1.8 220.1 1.8 68.7 1.4 4.5 0.2 3.0 0.1 62.71 1.7 47.1 1.1 150.7 1.3 27.5 1.9 172.8 1.6 99.4 1.4 4.4 0.2 3.2 0.3 63.9 48.3 161.9 32.8 219.4 84.6 4.2 3.2 2.8 3.8 3.2 2.2 1.6 1.4 0.2 0.3 62.9 49.7 439.7 87.5 323.8 113.8 4.4 3.6 Control 1.9 71.7 2.1 1.1 49.4 2.4 1.4 197.8 1.1 1.0 39.9 1.8 2.4 221.2 2.0 1.6 88.7 1.5 0.4 4.1 0.2 0.3 3.1 0.1

wastes such as urea, uric acid, creatinine and ions. During renal disease, the concentrations of these metabolites increase in blood. In this study, the levels of uric acid and creatinine did not appear to increase in any of the groups. This indicates the absence of any signicant kidney damage. The extract C lowered the elevated SGOT levels in diabetics by 21% with no effects in controls while glibenclamide lowered the elevated SGOT levels by 46% also with no effect in controls. SGPT levels are lowered by 14% in diabetics and 22% in controls by extract C and 12% in diabetics by glibenclamide with no effect in controls. Thus both extract C and glibenclamide reduced the increased levels of SGOT and SGPT observed in untreated diabetics in the present study. This might suggest the protective action of the extract and glibenclamide in reversing any organ damage due to induction of experimental diabetics that is manifested by elevations in the levels of SGOT and SGPT (Table 3). At the end of a 30-day treatment period, sections of liver and kidney showed no changes in the histology of the tissues. Oral feeding of extract powder C as well as glibenclamide for a period of 30 days also showed no changes in histology. Liver parenchymal cells showed normal architecture in all the groups. Hepatic lobules with portal tract and central vein with hepatic sinusoids radially arranged around the central vein were visible. The hepatic veins were normal in size and portal tract showed a collection of lymphocytes. No focal lesions were observed.

In the kidney, normal structure of Bowmans capsule was seen. The basement membrane appeared normal with no thickening. Normal tubular structures were also seen. No hemorrhagic or necrotic area could be detected. The histology of liver and kidney thus appeared similar and normal in all the groups. The lack of histological changes to kidneys is supported by absence of elevation in uric acid and creatine levels in urine. Though these results are indicative of a lack of active toxicity of the drug and are encouraging, it is essential to carry out studies on chronic toxicity.

4. Conclusion A systematic study of three extracts from Momordica charantia indicates that a water extract powder at doses as low as 20 mg/kg body weight can reverse alloxan induced hyperglycemia in rats with no toxicity to liver and kidneys up to a period of 4 weeks. Higher doses may not only be ineffective but may also cause toxicity. Thus Kulkarni and Gaitonde (1962) failed to observe any hypoglycemic response in rats fed bitter melon at 500 mg/kg body weight. The extract ameliorated loss in body weight in diabetic animals. The advantage of this natural preparation lies in its ability not only to control hyperglycemia at low dosages but can also be taken for longer periods. Thus bitter melon, a widely consumed vegetable could be safely prescribed to diabetic patients on

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a long term basis, which may also delay the onset of secondary complications.

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