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Shankaraiah Pulipaka et al. / Journal of Pharmacy Research 2011,4(12),4593-4595
Research Article
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Antidiabetic activity of methonolic extract of Operculina turpethum (l.)
Silva manso stem in streptozotocin induced diabetic rats
Shankaraiah Pulipaka1*, Challa Srinivas Reddy 2, Ravindra Babu. P3, SampathKumar.Ch1
1Trinity College of Pharmaceutical Sciences, Peddapally, Karimnagar, A.P, India, 505 172
2Vaagdevi College of Pharmacy, Ramnagar, Hanmakonda, Warangal, A.P India, 506001
3KVSR Siddhartha College of Pharmaceutical Sciences, Vijayawada, A. P, India, 520 010
Received on:20-08-2011; Revised on: 15-09-2011; Accepted on:10-11-2011
ABSTRACT
Diabetes mellitus (DM) consists of a group of syndromes characterized by hyperglycemia. the prevalence of both type 1 and type 2 DM is increasing worldwide,the prevalence of type 2 is rising much more rapidly because of increasing obesity and reduced activity levels as counties become more industrialized. Theantidiabetic potential of the methanolic extract of Operculina turpethum stem (MEOTS) (Convolvulaceae), a medicinal plant widely used in the traditionalAyurveda and Siddha systems of medicine for the treatment of diabetes mellitus was evaluated in the Streptozotocin (STZ) - induced type 2 diabetic models. Thedoses of 50 mg/kg and 100 mg/kg of MEOTS were administered to normal, glucose loaded and experimental diabetic rats for 21 days. Significant (p< 0.05) reductionin fasting blood glucose levels were observed in the normal rats at 3hr as well as in the treated diabetic animals at 21 days Significant results were observed in theestimated parameters, thereby justifying the use of the plant in the indigenous system of medicine.
Key words: Type 2 diabetes, STZ, Operculina turpethum , Glibenclamide
INTRODUCTION
Diabetes mellitus (DM) consists of a group of syndromes characterized by
Collection of plant material
hyperglycemia; altered metabolism of lipids, carbohydrates, and proteins; and Operculina turpethum (L.)Silva Manso plant material collected from local areas an increased risk of complications from vascular disease1. The worldwide preva- of Vijayawada, A.P. Its parts were botanically authenticated by Prof. S.V. Raju, lence of DM has raisen dramatically over the past two decades, from an esti- Taxonomist, Department of Botany, Kakatiya University, Warangal, A.P, and mated 30 million cases in 1985 to 177 million in 2000. Based on the current India. The herbarium was maintained in the Department of Pharmacognosy and trends > 360 million individuals will have diabetes by the year 20302.
phytochemistry, Vaagdevi College of pharmacy, Hanamkonda. Operculinaturpethum stem was washed under tap water and were efficiently dried under Operculina turpethum is a perennial with milky juice belongs to family shade for about one week and protected from deterioration. The shade dried Convolvulaceae. This plant was widespread in old tropics from E. Africa to N.
stem was grinded made into powder with the help of a laboratory mixer. These Australia, this plant common in Godavari, Andhra Pradesh, India. It is widely were efficiently dried under shade for about one week and protected from dete- distributed in tropical Africa and Asia. In India it is found in damp and it occurs rioration and then grinded and made into powder.
almost throughout India up to an altitude of about 1000 m. It is some timesgrown in gardens for its beautiful flowers. It is rare on open sandy soils. It is Preparation of extract
The chemical compounds were extracted from the stem using successive solventextraction process (soxhlation apparatus). The stem powder (100 g) was ex- Traditionally, Operculina turpethum root is prescribed in the treatment of tracted with methanol for 6 hours. After completion of soxhlation process the snake bite (sushruta and vrindamadhava) and scorpion sting (sushruta), but it is liquid extract was collected and concentrated under reduced pressure below 500C, not an antidote to either snake-venom (Mhaskar and Caius) or scorpion-venom until a soft mass obtained it was dried and kept in a desiccator.
(Caius and Mhaskar). In constipation, it is an effective laxative. It is used inperiodic fevers and in the treatment of anemia accompanied by spenomegaly. It Preparation of standard drug
is also used to relieve flatulence and colic and in the treatment of obesity to Glibenclamide was suspended in 0.5% Sodium Carboxy Methyl Cellulose (CMC).
decrease fat. It is used to treat dropsy, dyspepsia with constipation and flatu- 5 mg/kg of glibenclamide was administered to each rat in standard group.
lence, gout and rheumatism, and other inflammations4. In the present study,
stem methanolic extract was used to study hypoglycemic activity in normal and
Experimental design
Antidiabetic activity of MEOTS was assessed in normal, glucose loaded and STZ- induced diabetic rats. In all studies, the animals were fasted overnight for 16 h MATERIALS AND METHODS
with free access to water throughout the duration of the experiment.
Chemicals
Animals used
Streptozotocin was purchased from Sigma Aldrich chemicals Pvt. Ltd. USA.
Experiments were performed with male wistar rats procured from Mahaveera Glibenclamide was obtained as a gift sample from Sanofi Aventis India Ltd.
enterprises (Hyderabad, A.P., India), weighing between 180 - 220 g. The animals Glucometer (Bayer Health Care, Japan) purchased from local pharmacy. All were housed in individual polypropylene cages under standard laboratory condi- other chemicals and reagents used were of analytical grade.
tions of light, temperature (22 ± 1 oC) and relative humidity for at least one weekbefore the beginning of experiment, to adjust to the new environment and toovercome stress possibly incurred during transit. Animals were given standard rat *Corresponding author.
pellets and drinking water ad libitum. The animals were fasted 12 hours before Shankaraiah Pulipaka
the conduct of experiment and during the experiment they were withdrawn from Trinity College of Pharmaceutical Sciences,
food and water. The experiments were planned after the approval of Institu- Peddapally,
tional Animal Ethical Committee (IEAC).
Karimnagar, A.P, Pin: 505 172,India
Tel.: + 91-9885589543
Evaluation of MEOTS on normal healthy rats5
E-mail:[email protected]
At the end of the fasting period, taken as zero time (0 h), blood was withdrawn Journal of Pharmacy Research Vol.4.Issue 12.December 2011
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Shankaraiah Pulipaka et al. / Journal of Pharmacy Research 2011,4(12),4593-4595
from the tail vein. Blood glucose was estimated with glucometer. The animals Effect of MEOTS on oral glucose tolerance in normal rats
were then randomly divided into four groups of six animals each. Group I served MEOTS, when administered 60 min. prior to glucose loading produced signifi- as control and received 0.5% sodium CMC. Groups II treated with glibenclamide cant reduction (P < 0.05) in the rise in blood glucose levels at 60 min. after (5 mg/kg), group III and IV received MEOTS orally at the dose of 50 and 100 glucose administration. MEOTS at doses of 50 and 100 mg/kg produced 14.13% mg/kg. Blood glucose levels were determined 1, 2, 3 and 4 h following treatment and 19.89% reduction in blood glucose respectively when compared to vehicle Evaluation of MEOTS on oral glucose tolerance test6
Table 2: Effect of MEOTS on oral glucose tolerance test
Healthy rats were divided into four groups of six animals each. Group I served as Time (Min) Group I Group II Group III Group IV
control received 0.5% sodium CMC. Groups II treated with glibenclamide (5 mg/kg), group III and IV received MEOTS orally at the dose of 50 and 100 mg/kg.
0 88.6 ± 6.8 92.1 ± 6.1 94.1 ± 4.5 93.5 ± 8.7 30 85.8 ± 3.3 91.7 ± 5.1 91.5 ± 6.5 90.1 ± 5.9 All the animals were given glucose (2 g/kg) 60 min after dosing. Blood samples 60 82.6 ± 9.8 88.9 ± 3.7 90.1 ± 8.4* 85.2 ± 8.1* were collected from the tail vein just prior to (0 h) and at 30, 60, 90 and 120 min 90 79.5 ± 10.4 76.5 ± 6.2 86.4 ± 6.5* 80.5 ± 5.7* after the glucose loading, and blood glucose levels were estimated by glucometer.
120 84.5 ± 4.2 72.8 ± 4.2 80.8 ± 5.3* 74.9 ± 9.8* * P < 0.05 when compared to glibenclamide treated group Evaluation of MEOTS in STZ - induced diabetic rats7
Experimental diabetes was induced by single intraperitoneal injection of 55 mg/
kg of STZ, freshly dissolved in cold citrate buffer, pH 4.5. Control animals
received only citrate buffer. After 5 days of STZ injection, animals with fasting
blood glucose above 300 mg/dl were considered as diabetic and included in the
study. The animals were randomly divided into four groups of six animals each
and received the following treatments: Group I received 0.5% sodium CMC,
group II received glibenclamide (5 mg/kg), group III and IV received MEOTS
orally at the dose of 50 and 100 mg/kg. The freshly prepared solutions were
orally administered daily for 21 days. Blood glucose analysis was done weekly on
overnight fasted animals using glucometer. All data expressed as mean + S.D.
Statistical analysis was performed by ANOVA.
Effect of MEOTS on normoglycemic rats
Results of the effect of graded doses of MEOTS on blood glucose level of normal
healthy rats were presented in table 1 and figure 1. MEOTS produced peak
hypoglycemia at 3 h. Dose dependent blood glucose reduction was observed in
Fig 2: Table 2: Effect of MEOTS on oral glucose tolerance test
animals treated with 50 and 100 mg/kg (12.73% and 20.62%, respectively).
Effect of MEOTS on STZ - induced diabetic rats
Blood glucose levels were restored in all treatment groups by 4h.
The effect of repeated oral administration of stem methanolic extract on bloodglucose levels in STZ-diabetic rats were presented in table 3 and figure 3. MEOTS, Table 1: Effect of MEOTS on serum glucose level in normal rats
administered at three different doses of 50, 100 mg/kg to STZ-treated diabetic Time (hr) Group I Group II Group III Group IV
rats caused significant (P < 0.001) reduction of blood glucose levels which wasrelated to dose and duration of treatment. Maximum reduction was observed on 0 76.5 ± 3.3 74.6 ± 2.3 78.5 ± 2.6 76.2 ± 1.9 1 74.5 ± 2.1 72.1 ± 3.8 76.2 ± 4.5 73.8 ± 2.5 day 21 (30.5% and 43.66%, respectively). MEOTS 100 mg/kg exhibited maxi- 2 73.7 ± 3.5 65.2 ± 3.2 74.8 ± 3.2* 70.7 ± 3.4* mum glucose lowering effect in diabetic rats compared to the other dose.
3 72.3 ± 3.6 60.6 ± 4.5 68.5 ± 2.6** 60.4 ± 5.4** 4 73.6 ± 2.4 77.5 ± 2.9 76.3 ± 3.5 77.5 ± 4.3 Glibenclamide exhibited a 47.28% reduction in blood glucose levels at the end of * P < 0.01 when compared to glibenclamide treated group the study when compared to diabetic control.
* * P < 0.001 when compared to glibenclamide treated group Table 3: Effect of MEOTS on serum glucose level in STZ induced dia-
betic rats
Day Group I Group II Group III Group IV
1 271.1±11.3 242.9±10.6 218.6±6.8 222.6±18.8 7 269.0±6.8 207.5±11.6 157.0±2.3** 194.4±16.0** 14 263.8±12.7 181.4±4.8 149.4±8.4** 144.5±10.9** 21 265.3±12.5 128.0±6.4 151.9±7.9** 125.4±8.6** * * P < 0.001 when compared to glibenclamide treated group Fig 1: Effect of MEOTS on serum glucose level in normal rats
Fig 3: Effect of MEOTS on serum glucose level in STZ induced diabetic rats
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Shankaraiah Pulipaka et al. / Journal of Pharmacy Research 2011,4(12),4593-4595
DISCUSSION
Pharmacological Basis of Therapeutics, 11th ed. McGraw Hill Publishing. 2006; This study was undertaken to evaluate the hypoglycemic activity of MEOTS in normal, glucose loaded and STZ - induced diabetic rats. In normoglycemic rats, Harisons principles of internal medicine, 17th edition. 2007; 2275 – 77 MEOTS showed dose dependent hypoglycemic effect at 3 h. From oral glucose John A, Parrotta. Healing plants of Penisular India. USDA Forest service, Interna- tolerance test (OGTT) it could be concluded that dose of 100 mg/kg showed the tional Institute of Tropical Forestry; 239-251.
Suresh kumar S.V, Sujatha C, Shymala. Protective effect of root extract of Operculina maximum improvement in glucose tolerance. STZ significantly induced hyper- turpethum Linn. Against Paracetamol induced Hepatotoxicity in rats. Indian Jour- glycemia accompanied by hypoinsulinemia. Oral administration of MEOTS for nal of Pharmaceutical Sciences. 2006; 68, 32- 35.
21 days caused a significant decrease in blood glucose levels. The possible mecha- Kar, D, Maharana L, Pattnaik, S, Dash G. Studies on hypoglycaemic activity of nism by which MEOTS mediated its antidiabetic effect could be by potentiation Solanum xanthocarpum Schrad. & Wendl. Fruit extract in rats. Journal of of pancreatic secretion of insulin from existing ß –cells of islets, as was evident Ethnopharmacology. 2006; 108, 251–256.
by the significant increase in the level of insulin in the extract treated animals.
Prakasam A., Sethupathy S., Pugalendi, K. Effect of Casearia esculenta root extract onblood glucose and plasma antioxidant status in streptozotocin diabetic rats. Polish The hypoglycemic activity of MEOTS was compared with glibenclamide, a Journal of Pharmacology. 2003; 55, 43–49.
standard hypoglycemic drug. From the results of the present study, it may be Arulselvan, P., Subramanian, S. Beneficial effects of Murraya koenigii leaves on an- suggested that the mechanism of action of MEOTS may be similar to tioxidant defense system and ultra structural changes of pancreatic -cells in experi- mental diabetes in rats. Chemico-Biological Interactions. 2007; 165, 155–164.
Kirtikar, J., Basu, B. Ficus religiosa. In: Blatter, E., Caius, J., Mhaskar, K. (Eds.), CONCLUSION
Indian Medicinal Plants, 2nd ed. Oriental Enterprises. 2001; 3200–3202.
From this study, we can conclude that MEOTS has beneficial effects on blood Vogel H.G, Vogel W.H., Drug Discovery and evaluation – pharmacological assays, glucose level. It has the potential to impart therapeutic effect in diabetes.
Szkudelski T. The mechanism of alloxan and Streptozotocin action in ß-cells of the ratpancreas, Physiololgicl Research. 2001; 50, p.p 536.
REFERENCES
Rucha Pandit, Ashish Phadke, Aarti Jagtap. Antidiabetic effect of Ficus religiosa Davis S. Insulin, oral hypoglycemic agents and the pharmacology of the endocrine extract in streptozotocin-induced diabetic rats. Journal of Ethnopharmacology. 2010; pancreas. In Brunton, LL, Lazo JS, Parker KL. eds. Goodman and Gilman’s The Source of support: Nil, Conflict of interest: None Declared
Journal of Pharmacy Research Vol.4.Issue 12.December 2011
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Source: http://kaubic.in/DIACAN/Operculina_turpethum_Ref1.pdf