hypoglycaemic effect of melothria heterophylla in streptozotocin-induced...

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Introduction

Diabetes mellitus, both insulin-dependent diabetes mellitus (IDDM) and nonIDDM type, is a common and serious disorder throughout the world (Keen, 1986; Harris et al., 1987). This metabolic disorder often leads to physical disability arising from the vascular compli-cations of coronary artery disease and cerebrovascular disease, resulting in renal failure, blindness, and limb amputation in addition to neurological complications and premature death (Weidmann et al., 1993; Cho et al., 2005). Treatment of diabetes mellitus by insulin and oral hypoglycemic drugs fails to prevent these complications

in many patients, indicating that additional alternative treatment could be helpful. Plant derived drugs are gain-ing popularity in the treatment of diabetic mellitus (Pari, 1999). The major advantages of herbal medicine seem to be their efficacy, low incidence of side effects. Melothria heterophylla (Lour.) Cogn. (Cucurbitaceae), popularly known as kudari, is a scandent herb with tuberous roots found throughout India ascending up to 2100 m in the hills. It is reported to be useful for stimulant, invigorat-ing, diabetes, cuts, fever, anti-inflammatory and purga-tive properties (Kirtikar & Basu, 2000; Anonymous, 1962; Pant & Samant, 2010) and antioxidant (Mondal et al.,

research artIcle

Hypoglycaemic effect of Melothria heterophylla in streptozotocin-induced diabetic rats

Arijit Mondal1, Tapan Kumar Maity1, and Dilipkumar Pal2

1Department of Pharmaceutical Technology, Jadavpur University, Kolkata - 700 032, India and 2School of Pharmaceutical Sciences, IFTM University, Moradabad-244 001, India

abstractContext: In the Indian traditional system of medicine, Melothria heterophylla (Lour.) Cogn., (Cucurbitaceae) is prescribed for the treatment of diabetes mellitus.Objective: In the present study, the antidiabetic effect of ethanol extract of Melothria heterophylla (EEMH), and its active isolated constituents were investigated in streptozotocin (STZ)-induced diabetic Swiss albino rats.Method: Successive Soxhlet extraction of the dried total aerial parts with petroleum ether for defatting and then with ethanol (95%) to obtain ethanol extract, which was concentrated under reduced pressure. Hyperglycemia was induced in rats by STZ (50 mg/kg, body weight). Twenty-four hours after STZ induction, respective groups of diabetic rats received EEMH (200 and 400 mg/kg, body weight), gallic acid (GA) (2 and 4 mg/kg, body weight), and rutin (RU) (2 and 4 mg/kg, body weight), respectively, orally daily for 15 days. Glibenclamide (0.5 mg/kg, orally) served as reference. Blood glucose levels and change in body weight were measured on every 5th day during 15 days of treatment. Biochemical parameters, viz., serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase (ALP) and serum insulin, were measured.Results: EEMH and its active constituents significantly (p < 0.01) normalized blood glucose levels and serum biochemical parameters as compared to those of STZ controls. Both GA (4 mg/kg) and RU (4 mg/kg) exhibited maximum glucose lowering effect (69.1 and 66.7%, respectively) in diabetic rats compared to the other dose (2 mg/kg) at the end of the study. EEMH, gallic acid and RU also showed significant increase in serum insulin, and body weight of STZ-induced diabetic rats.Conclusion: Therefore, ethanol extract of Melothria heterophylla, GA and RU demonstrated remarkable antidiabetic activity in STZ-induced diabetic rats.Keywords: Diabetes mellitus, gallic acid, rutin

Address for Correspondence: Arijit Mondal, Department of Pharmaceutical Technology, Jadavpur University, Kolkata, West Bengal-700 032, India. E-mail: [email protected]

(Received 05 September 2011; revised 10 November 2011; accepted 03 January 2012)

Pharmaceutical Biology, 2012; 50(9): 1151–1156© 2012 Informa Healthcare USA, Inc.ISSN 1388-0209 print/ISSN 1744-5116 onlineDOI: 10.3109/13880209.2012.661742

Pharmaceutical Biology

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© 2012 Informa Healthcare USA, Inc.

10.3109/13880209.2012.661742

2012

A. Mondal et al.

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1152 A. Mondal et al.


2009). However, there is no scientific evidence to sup-port the antidiabetic effect of Melothria heterophylla. Hence, the objective of this study was to evaluate the antidiabetic activity of ethanol extracts of the aerial parts of M. heterophylla and its active isolated constituents in streptozotocin (STZ)-induced diabetic rats.

Material and methods

Plant materialThe aerial parts of Melothria heterophylla were col-lected from young matured plants during August and September 2010, from the rural belt of Mayurbhanj district, Odisha, India and identified by Dr. P. Venu, tax-onomist of Botanical Survey of India, Howrah, India. A voucher specimen (CNH/I-I (65)2006/Tech.II/1661) was deposited in the Department of Pharmaceutical Technology, Jadavpur University, Kolkata, India. The col-lected plant material was washed, shade-dried, and then milled to course powder by a mechanical grinder for fur-ther studies.

Extraction and isolationThe powdered plant material (2 kg) was defatted with petroleum ether (60–80°C) and then extracted with 3L of ethanol (95%) in a Soxhlet apparatus. The solvent was then removed under reduced pressure, to obtain petroleum ether (Melothria heterophylla (PEMH), yield 4.38%) and ethanol (EEMH, yield 10.1%) extract, respectively. The eth-anol extract was partitioned successively between ethyl acetate-water system and then between n-butanol-water system (3×1L). The respective solvents were removed similarly under reduced pressure, which pro-duced ethyl acetate fraction (EAF) (120 g) and n-butanol fraction (NBF) (60 g). Both the fractions were evaluated for antidiabetic activity in STZ-induced diabetic rats. EAF was found to be more potent than NBF. Hence, EAF was further exploited for isolation, which led to the iso-lation of gallic acid (GA), a phenolic acid and rutin (RU), a flavonoid glycoside. GA was isolated as a white colored compound and RU as a yellowish amorphous powder, which were characterized as GA and RU based on their melting point and spectroscopic (IR, 1H, 13C Nuclear magnetic resonance (NMR) and Mass Spectrometry (MS)) data (Latha & Daisy, 2011; Abdullah et al., 2008).

AnimalsMale Swiss albino rat weighing 150–250 g were used for the present investigation. They were housed in clean polypropylene cages and were fed with standard pel-let diet (Hindustan Lever, Kolkata, India) and water ad libitum. The animals were acclimatized to laboratory condition (temperature 25 ± 2°C) with dark/light cycle (14/10 h) for one week before the start of an experiment. All procedures were approved by Jadavpur University Animal Ethical Committee (CPCSEA/ORG/CH/2006/Reg.No.95), Kolkata, India.

Drugs and chemicalsSTZ was obtained from Spectrochem Pvt. Ltd, Mumbai, India; glibenclamide was obtained from Hoechst, India. All other reagents used were of analytical grade and were obtained commercially.

Acute toxicity testThe animals were divided into five groups (n = 6). The EEMH suspension was administrated orally in increasing dose up to 2000 mg/kg, b.w (Litchfield & Wilcoxon, 1949). The rats were observed continuously for 2 h for behav-ioral, neurological and autonomic profiles and after 24 and 72 h for any lethality (Turner, 1965).

Induction of experimental diabetesDiabetes was induced by a single intraperitoneal injec-tion of freshly prepared STZ (50 mg/kg, b.w) in ice cold citrate buffer (0.1 M, pH 4.5) to overnight-fasted rats (Yoruk et al., 2003; Kanter et al., 2003). Diabetes was identified by measuring glucose levels 48 h after injection of STZ. Only rats with glucose levels greater than 300 mg/dL were used in experiments.

Oral Glucose Tolerance Test in normal and STZ-diabetic ratsThe oral glucose tolerance test (OGTT) was performed for two different doses of the EEMH (at predetermined therapeutic doses), and its isolated compounds (GA and RU) in normal and STZ-induced diabetic rat model. Four days after diabetes induction, the OGTT was performed by feeding glucose in the form of a solution through orally to STZ-induced diabetic rats fasted for 18 h. The animals were divided into nine groups, each having six animals. Group-I served normal control and received normal saline only; the animals of group II were administered STZ only; group III-IV were treated with EEMH 200 and 400 mg/kg, b.w., p.o; group V-VI with GA 2 and 4 mg/kg, b.w., p.o; group VII-VIII with RU 2 and 4 mg/kg, b.w., p.o and group IX was treated with glib-enclamide (a known hypoglycemic agent) 0.5 mg/kg (Das et al., 2011), respectively. One hour later, glucose (2 g/kg) was administrated. EEMH and its isolated compounds were administered throughout 14 days consecutively. The blood glucose level of each animal was monitored on 0, 5th, 10th and 15th days after the administration of the test samples.

Estimation of blood glucoseBlood glucose levels from the tail vein were measured after glucose administration using one touch system Accu-Check Active Glucometer (Roche Diagnostics India Pvt. Ltd, India).

Biochemical estimationsDifferent biochemical parameters such as serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT) and alkaline phosphatase (ALP) were determined by using commercially available kits (Span Diagnostic Limited, Surat, India) (Reitman & Frankel, 1957).

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Hypoglycaemic effect of Melothria heterophylla 1153


Serum insulin estimation was done spectrophotometrically using standard kits (Span Diagnostic Limited, Surat, India).

Statistical analysisData were statistically evaluated using one-way analysis of variance (ANOVA), followed by post hoc Dunnett’s test using version 10 of SPSS computer software. The values are expressed as mean ± SD and p values less than 0.01 was considered as significant.

results

Acute toxicity studiesAcute toxicity studies revealed the non-toxic nature of the EEMH. There was no lethality or toxic reaction found at any doses selected until the end of the study period.

Acute administration of GA even at a dose as high as 5 g/kg body weight did not produce any signs of toxicity or mortality (Rajalakshmi et al., 2001). RU is also reported to be safe as well (Palmer et al., 2002).

Effect of EEMH, GA and RU on fasting blood glucose and body weight in STZ-induced diabetic ratsAfter 24 h of treatment with STZ, the fasting blood-glucose level was significantly changed in the range of

300–350 mg/dL (Table 1). Treatment of EEMH led to a dose-dependent fall in blood sugar levels by 4.9–70.1%. The initial antidiabetic activity was observed on the 5th day. The observed effect with isolated compounds was more significant (p < 0.01) pronounced than that of the EEMH. Both GA and RU (4 mg/kg) exhibited maximum glucose lowering effect (69.1 and 66.7%, respectively) in diabetic rats compared to the other dose (2 mg/kg) at the end of the study. Glibenclamide significantly (p < 0.01) exhibited a 61.70% reduction in blood-glucose level at the end of the study when compared to diabetic control.

STZ produced significant (p < 0.01) loss in body weight as compared to normal animals during the study. Diabetic control continued to loose weight till the end of the study while EEMH, GA and RU showed significant improvement (p < 0.01) in body weight compared to diabetic control (Table 2).

Effect of EEMH, GA and RU on serum insulin in STZ-induced diabetic ratsSTZ caused a significant (p < 0.01) decrease in serum insu-lin. Administration of EEMH and its isolated compounds caused significant (p < 0.01) increase in insulin levels at the end of the study period. Amongst the drug treated

Table 1. Effect of EEMH and GA and RU on the glucose level in STZ-induced diabetic rats.

GroupsSerum glucose levels (mg/dL)

0 day 5th day 10th day 15th dayNormal (0.9% NaCl) 102.30 ± 0.87 101.60 ± 0.99 100.20 ± 1.01 94.46 ± 0.68STZ control 322.40 ± 1.42a* 326.0 ± 0.97a* 331.90 ± 1.27a* 326.20 ± 0.70a*TZ+ EEMH (200 mg/kg) 423.80 ± 1.94b* 310.10 ± 1.38b* 230.80 ± 1.30b* 145.30 ± 0.95b*STZ+ EEMH (400 mg/kg) 480.60 ± 0.65b* 202.20 ± 0.96b* 134.50 ± 1.35b* 97.49 ± 0.76b*STZ+ GA (2 mg/kg) 367.60 ± 1.13b* 189.10 ± 1.26b* 144.6 ± 1.05b* 108.1 ± 0.89b*STZ+ GA (4 mg/kg) 358.90 ± 2.07b* 139.20 ± 1.21b* 110.20 ± 1.18b* 100.70 ± 1.73b*STZ+ RU (2 mg/kg) 410.20 ± 0.69b* 228.90 ± 1.47b* 157.70 ± 1.36b* 114.30 ± 1.01b*STZ+ RU (4 mg/kg) 399.0 ± 1.42b* 200.30 ± 1.08b* 142.70 ± 1.32b* 108.70 ± 1.30b*STZ+ Glibenclamide (0.5 mg/kg) 357.20 ± 0.98b* 227.30 ± 2.47b* 158.3 ± 1.45b* 125.00 ± 1.03b*(Values are mean ± SD, n = 6). STZ (50 mg/kg., b.w) was injected to control and all other drugs treated groups; a STZ induced diabetic group vs normal group, *p < 0.01; bdrug treated group vs STZ induced diabetic group, *p < 0.01.EEMH, ethanol extract of Melothria heterophylla; GA, gallic acid; STZ, streptozotocin; RU, rutin.

Table2. The effect on body weight changes during 14 days treatment of EEMH, GA and RU.

Groups Dose (mg/kg., b.w.)Body weight (g/day)

0 day 5th day 10th day 15th dayNormal --- 211.2 ± 1.15 219.2 ± 2.27 226.4 ± 0.79 231.9 ± 2.03STZ control --- 181.6 ± 1.00 175.0 ± 1.44*a 164.4 ± 1.40*a 158.5 ± 1.21*a

STZ + EEMH 200 181.6 ± 1.72 183.8 ± 1.06*b 190.2 ± 1.14*b 200.4 ± 1.77*b

STZ + EEMH 400 185.0 ± 0.78 188.3 ± 1.19*b 195.6 ± 1.73*b 203.8 ± 1.14*b

STZ + GA 2 183.5 ± 0.96 186.1 ± 0.35*b 198.0 ± 1.11*b 210.5 ± 1.43*b

STZ + GA 4 184.1 ± 0.62 188.8 ± 0.80*b 199.5 ± 1.31*b 212.5 ± 1.73*b

STZ + RU 2 181.3 ± 0.86 183.6 ± 0.85*b 188.7 ± 1.75*b 197.1 ± 0.86*b

STZ + RU 4 182.4 ± 0.72 185.2 ± 0.79*b 192.4 ± 1.66*b 199.1 ± 0.97*b

STZ + Glibenclamide 0.5 181.3 ± 0.82 193.3 ± 3.26*b 202.4 ± 2.23*b 220.4 ± 0.65*b

(Values are mean ± SD, n = 6). STZ (50 mg/kg., b.w) was injected to control and all other drugs treated groups; a STZ induced diabetic group vs normal group, *p < 0.01; b treated group vs STZ induced diabetic group, *p < 0.01.EEMH, ethanol extract of Melothria heterophylla; GA, Gallic acid; STZ, streptozotocin; RU, rutin.

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groups, GA (4 mg/kg, b.w.) showed maximum increase which was comparable to glibenclamide (Table 3).

Biochemical enzymesTable 4 shows the effect of the EEMH on the serum glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and ALP levels in normal and diabetic rats. The results showed that serum GOT, GPT and ALP levels in diabetic groups increased, when compared with normal rats. The administration of the GA and rutin significantly (p < 0.01) decreased the serum GOT, GPT and ALP levels, when compared with control diabetic rats.

Discussion

STZ is broad-spectrum antibiotic from Streptomyces achromogenes. Since the finding, that STZ possesses diabetogenic properties mediated by pancreatic β-cell destruction; this compound has been widely used to induce diabetes in experimental animals. Evidence has

accumulated suggesting that STZ induces oxidative stress, which is caused by a relative overload of reactive oxygen species (ROS). Once, STZ enters inside the cell, it is spontaneously decomposed to form an isocyanate compound and a methyldiazohydroxide. Isocyanate compound and methyldiazohydroxide undergo intra molecular carboxylation and alkylation of cellular com-ponents respectively. The DNA damage of β cells of the pancreas, mainly by alkylation with carbonium ion pro-duced by methyldiazohydroxide (Weiss, 1982; Wilson & Leiter, 1990). It is well known that diabetes mellitus causes a disturbance in the uptake of glucose as well as glucose metabolism. Even the low dose of STZ produced an incomplete destruction of pancreatic β-cells even though the rats become permanently diabetic and induce disturbance in glucose metabolism (Aybar et al., 2001). The induction of diabetes was confirmed as reflected by the hyperglycemia, and body weight loss (Mi-Kyung et al., 2005; Cho et al., 2006a,b). The single high-dose STZ-induced diabetic rat is one of the animal models of human IDDM or type I diabetes mellitus. In this model, diabetes arises from irreversible destruction of the islet cells of the pancreas, causing degranulation or reduction of insulin secretion and leads to hyperglycemia. In this type I model of diabetes, insulin is markedly depleted, but not absent (Pushparaj et al., 2001). Although insulin has become one of the most important therapeutic agents known to medicine, there is a continuing effort to find insulin substitutes, secretagogues, or sensitizers from synthetic or plant sources for the treatment of diabetes. Phyto-constituents, such as saponins and flavonoids, play important roles as the hypoglycemic agents (Pushparaj et al., 2001; Hosseinzadeh et al., 2002). The isolated plant compounds such as GA and rutin from M. heterophylla are phenolic acid and flavonoid glycoside respectively, and the plants were used by traditional people for the treatment of diabetes. In the present study, the high dose of EEMH and its secondary metabolites had significantly reduced hyperglycemia in STZ diabetic rats.

Hyperglycemia is associated with the generation of ROS causing oxidative damage, particularly to the heart, kidney, eyes, nerves, liver, small and large vessels and

Table 3. Effect of EEMH, GA and RU on serum insulin of STZ-treated diabetic rats.

Groups

Dose (mg/kg.,

b.w.)

Serum insulin (µU/mL)

0 day 15th dayNormal --- 18.19 ± 0.19 18.65 ± 0.27STZ control --- 8.45 ± 0.18 07.23 ± 0.15*a

STZ + EEMH 200 8.34 ± 0.19 12.14 ± 0.41*b

STZ + EEMH 400 8.42 ± 0.34 16.21 ± 0.34*b

STZ + GA 2 8.39 ± 0.17 12.91 ± 0.60*b

STZ + GA 4 8.58 ± 0.28 17.43 ± 0.36*b

STZ + RU 2 8.24 ± 0.20 13.34 ± 0.33*b

STZ + RU 4 8.33 ± 0.07 15.98 ± 0.63*b

STZ + Glibenclamide

0.5 8.45 ± 0.25 18.16 ± 0.17*b

(Values are mean ± SD, n = 6). STZ (50 mg/kg., b.w) was injected to control and all other drugs treated groups; a STZ induced diabetic group vs normal group, *p < 0.01; btreated group vs STZ induced diabetic group, *p < 0.01.EEMH, ethanol extract of Melothria heterophylla; GA, gallic acid; STZ, streptozotocin; RU, rutin.

Table 4. Effect of ethanol extract of M. heterophylla, GA and RU on serum biomarkers in STZ-induced diabetic rats after 14 days treatment.Groups SGOT (IU/dL) SGPT (IU/dL) ALP (IU/dL)Normal (0.9% NaCl) 57.20 ± 1.38 61.06 ± 0.92 121.70 ± 1.49STZ control 146.10 ± 0.98a* 132.40 ± 0.56a* 245.50 ± 0.87a*STZ + EEMH (200 mg/kg) 92.15 ± 0.98 b* 87.01 ± 1.07 b* 181.70 ± 1.06 b*STZ + EEMH (400 mg/kg) 70.15 ±1.01 b* 76.05 ± 0.76 b* 155.00 ± 0.95 b*STZ + GA (2 mg/kg) 58.89 ± 0.55 b* 65.61 ±0.97 b* 134.40 ±1.33 b*STZ + GA (4 mg/kg) 55.36 ± 0.91 b* 60.67 ±0.92 b* 124.50 ± 0.92 b*

STZ + RU (2 mg/kg) 61.07 ± 0.89 b* 67.83 ± 0.86 b* 132.90 ± 0.77 b*STZ + RU (4 mg/kg) 59.07 ± 1.11 b* 63.92 ± 1.75 b* 128.90 ± 0.93 b*STZ + Glibenclamide (0.5 mg/kg) 63.63 ± 0.77 b* 72.11 ± 1.11 b* 133.50 ± 0.82 b*(Values are mean ± SD, n = 6). STZ (50 mg/kg., b.w) was injected to control and all other drugs treated groups; aSTZ induced diabetic group vs normal group, *p < 0.01; btreated group vs STZ induced diabetic group, *p < 0.01.EEMH, ethanol extract of Melothria heterophylla; GA, gallic acid; STZ, streptozotocin; RU, rutin.

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gastrointestinal system. The increased levels of plasma glucose in STZ-induced diabetic rats were lowered by the administration of EEMH, GA and rutin. The plasma glu-cose lowering activity was compared with glibenclamide, a standard hypoglycemic drug. The possible mechanism by which EEMH and its isolated compounds mediated their antidiabetic effect could be by potentiation of pancreatic secretion of insulin from existing β-cells of islets (Tian et al., 1998), as was evident by the significant increase in the level of insulin in the extract and isolated compounds treated animals. The hypoglycemic activity of EEMH, GA and RU was compared with glibenclamide. From the results of the present study, it may be suggested that the mechanism of action of FRAE may be similar to glibenclamide action.

The increase in the activities of serum GOT, GPT and ALP indicated that diabetes may be induced due to liver dysfunction in STZ-induced diabetic rats. Therefore, increase in the activities of GOT, GPT and ALP in plasma may be mainly due to the leakage of these enzymes from the liver cytosol into the blood stream (Fortson et al., 1985), which gives an indication on the hepatotoxic effect of STZ. On the other hand, independent treatment of the diabetic rats with GA and rutin caused the reduction in the activity of these enzymes in plasma compared to the mean values of the diabetic group and consequently, may alleviate liver damage caused by STZ-induced diabetes. These results are in agreement with those obtained by El-Demerdash et al. (2005) in rats. This investigation reveals the potential effect of GA and rutin as the hypoglycemic agent.

conclusion

In conclusion, it can be stated, that EEMH and its iso-lated compounds, GA and rutin, have marked beneficial effects, in reducing the elevated blood-glucose level of STZ-induced diabetic rats, justifying the folklore claim.

acknowledgement

The authors would like to thank University Grants com-mission (UGC), New Delhi, India, for providing financial assistance and Jadavpur University, Kolkata, India, for providing research facilities to the corresponding author.

Declaration of interest

The authors report no declarations of interest.

referencesAbdullah Y, Schneider B, Petersen M. (2008). Occurrence of rosmarinic

acid, chlorogenic acid and rutin in Marantaceae species. Phyt Letters, 1, 199–203.

Anonymous. (1962). The Wealth of India. New Delhi, India.: Publi-cations and Information Directorate, C.S.I.R, 6, 335.

Aybar MJ, Sánchez Riera AN, Grau A, Sánchez SS. (2001). Hypoglycemic effect of the water extract of Smallantus sonchifolius (yacon) leaves in normal and diabetic rats. J Ethnopharmacol, 74, 125–132.

Cho WC, Yue KK, Leung AW. (2005). An outline of diabetes mellitus and its treatments by traditional Chinese medicine and acupuncture. J Chin Med, 78, 29–37.

Cho WC, Chung WS, Lee SK, Leung AW, Cheng CH, Yue KK. (2006a). Ginsenoside Re of Panax ginseng possesses significant antioxidant and antihyperlipidemic efficacies in streptozotocin-induced diabetic rats. Eur J Pharmacol, 550, 173–179.

Cho WC, Yip TT, Chung WS, Lee SK, Leung AW, Cheng CH, Yue KK. (2006b). Altered expression of serum protein in ginsenoside Re-treated diabetic rats detected by SELDI-TOF MS. J Ethnopharmacol, 108, 272–279.

Das S, Bhattacharya S, Prasanna A, Suresh Kumar RB, Pramanik G, Haldar PK. (2011). Preclinical evaluation of antihyperglycemic activity of Clerodendron infortunatum leaf against streptozotocin-induced diabetic rats. Diabetes Ther, 2, 92–100.

El-Demerdash FM, Yousef MI, El-Naga NI. (2005). Biochemical study on the hypoglycemic effects of onion and garlic in alloxan-induced diabetic rats. Food Chem Toxicol, 43, 57–63.

Fortson WC, Tedesco FJ, Starnes EC, Shaw CT. (1985). Marked elevation of serum transaminase activity associated with extrahepatic biliary tract disease. J Clin Gastroenterol, 7, 502–505.

Harris MI, Hadden WC, Knowler WC, Bennett PH. (1987). Prevalence of diabetes and impaired glucose tolerance and plasma glucose levels in U.S. population aged 20-74 yr. Diabetes, 36, 523–534.

Hosseinzadeh H, Ramezani M, Danaei AR. (2002). Antihyperglycaemic effect and acute toxicity of Securigera Securidaca L. seed extracts in mice. Phytother Res, 16, 745–747.

Kanter M, Yoruk M, Koc A, Meral I, Karaca T. (2003). Effects of cadmium exposure on morphological aspects of pancreas, weights of fetus and placenta in streptozotocin-induced diabetic pregnant rats. Biol Trace Elem Res, 93, 189–200.

Keen H. (1986). What’s in a name? IDDM/NIDDM, type 1/type 2. Diabet Med, 3, 11–12.

Kirtikar KR, Basu BD. (2000). Indian Medicinal Plants. 3rd edn. Delhi, India: Sri Satguru Publication, 5, 1618.

Latha RC, Daisy P. (2011). Insulin-secretagogue, antihyperlipidemic and other protective effects of gallic acid isolated from Terminalia bellerica Roxb. in streptozotocin-induced diabetic rats. Chem Biol Interact, 189, 112–118.

Litchfield JT Jr, Wilcoxon F. (1949). A simplified method of evaluating dose-effect experiments. J Pharmacol Exp Ther, 96, 99–113.

Mi-Kyung L, Myung-Joo K, Soo-Yeul C, Sun Ae P, Kwan-Kyu P, Un Ju J, Hae-Mo P, Myung-Sook C. (2005). Hypoglycemic effect of Du-zhong (Eucommia ulmoides Oliv.) leaves in streptozotocin-induced diabetic rats. Diabetes Res Clin Pract, 67, 22–28.

Mondal A, Maity TK, Pal DK, Sannigrahi S. (2009). (In vitro antioxidant activity of the roots of Melothria heterophylla Lour Cogn. 2009). Pharmacologyonline, 2, 499–509.

Palmer H, Ohta M, Watanabe M, Suzuki T. (2002). Oxidative stress-induced cellular damage caused by UV and methyl viologen in Euglena gracilis and its suppression with rutin. J Photochem Photobiol B, 67, 116–129.

Pant S, Samant SS. (2010). Ethanobotanical observations in the Mornaula Reserve Forest of Kumoun, West Himalaya India. Ethnobotanical Leaflets, 14, 193–217.

Pari L, Maheswari JU. (1999). Hypoglycaemic effect of Musa sapientum L. in alloxan-induced diabetic rats. J Ethnopharmacol, 68, 321–325.

Pushparaj NP, Tan HKB, Tan HC . (2001). The mechanism of hypoglycemic action of the semi-purified fractions of Averrhoa bilimbi in streptozotocin diabetic rats. Life Sciences, 70, 535–547.

Rajalakshmi K, Devaraj H, Niranjali Devaraj S. (2001). Assessment of the no-observed-adverse-effect level (NOAEL) of gallic acid in mice. Food Chem Toxicol, 39, 919–922.

Reitman S, Frankel S. (1957). A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol, 28, 56–63.

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al B

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oade

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ahea

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nive

rsity

of

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ches

ter

on 1

1/11

/14

For

pers

onal

use

onl

y.



Tian YA, Johnson G, Ashcroft SJ. (1998). Sulfonylureas enhance exocytosis from pancreatic beta-cells by a mechanism that does not involve direct activation of protein kinase C. Diabetes, 47, 1722–1726.

Turner MA. (1965). Screening Methods in Pharmacology. New York: Academic Press.

Weidmann P, Boehlen LM, de Courten M. (1993). Pathogenesis and treatment of hypertension associated with diabetes mellitus. Am Heart J, 125, 1498–1513.

Weiss RB. (1982). Streptozocin: A review of its pharmacology, efficacy, and toxicity. Cancer Treat Rep, 66, 427–438.

Wilson GL, Leiter EH. (1990). Streptozotocin interactions with pancreatic beta cells and the induction of insulin-dependent diabetes. Curr Top Microbiol Immunol, 156, 27–54.

Yoruk M, Kanter M, Meral I, Agaoglu Z. (2003). Localization of glycogen in the placenta and fetal and maternal livers of cadmium-exposed diabetic pregnant rats. Biol Trace Elem Res, 96, 217–226.

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