INTRODUCTION:

Indian traditional medicine is based on various systems including Ayurveda, Siddha and Unani. Diabetes mellitus (DM) is a multi-factorial disease and it is of two types, type1 and type 2. Type -2 diabetes is a major chronic disorder and it is a major public health problem in the developed as well as developing countries caused by partial or complete insulin deficiency, resulting in hyperglycemia leading to acute and chronic complication¹. Synthetic drugs are likely to give serious side effects in addition they are not suitable for intake during conditions like pregnancy². There are various herbal remedies for diabetes described in Ayurveda (The ancient healthcare system of India). These herbs are effective in healing the diabetic ulcers and keeping the sugar levels under control without causing any side effects. The herbs can also be used by females to restore libido, fatigue, general weakness and pain in the calf muscles due to diabetic mellitus as well as high sugar levels and it is useful in regulating the menstrual cycle. The flower and flower buds can be used beneficially as pessaries to check excessive menstrual flow³. Cassia auriculata L is generally found in warm and moist climate.

The seeds of tanner’s cassia find their application in purulent opthalmia i.e., inflammation of the eye or conjunctiva. They should be finely powdered and blown into the affected eyes⁴. The bark of the plant is useful in checking secretion or haemorrhage. They also restore the disordered processes of nutrition. Its leaves and petals are both mildly astringent in taste⁵. It also checks the flow of extra amount of urine and helps in absorption of required amount of fluids in the kidneys and intestines⁶. It is used to be effective in numerous cases like, diabetes, conjunctivitis, aching throat, troubled menstruation, opthalmia, also arresting bleeding. Even diseases like diarrhea and dysentery can be cured by the plant⁷.

MATERIALS AND METHODS:

Streptazotocin, nicotinamide and glibenclamide (Sigma Aldrich, Missouri, U.S.A), Hydrogen peroxide (Qualikems Fine Chemicals, India), Glucometer kit (Accue check) were procured from local market. All the solvents and chemicals were procured from S.D fine chemicals, Mumbai, India and they were analytical grade quality.

COLLECTION OF PLANT MATERIAL

Dried flower of Cassia auriculata L. were collected from Kattangur Village, Nalgonda, Andra Pradesh, India. The plant was authenticated by Dr. Sai Reddy, Department of Botany A.P.S.W.R College, Osmania University, Hyderabad, A.P., INDIA.

PREPARATION OF EXTRACT

The flowers of Cassia auriculata L. were collected and shade dried at room temperature the dried flowers were subjected to size reduction to a coarse powder by using the dry grinding mixer, passed through the sieve (50 mesh). The powder (80 gram) was extracted with 5 liters of ethanol overly for 7 days by method of cold maceration at room temperature occasionally mixing the contents for every 6 hours. The container was closed with lid to prevent the evaporation of ethanol. After 7 days the contents are filtered and the filtrate is evaporated, air-dried and kept in desiccator. The yield of ethanolic extract obtained was about 15 grams i.e. 18.75 %. The concentrated residue was stored in air tight container in refrigerator (2 – 8 ⁰C) for use in further experimentations. The suspension of extract was prepared freshly by using 1 % CMC^9,10.

EXPERIMENTAL ANIMALS

Male albino (Wistar Strain) rats (200 - 250 g) were procured from the National Institute of Nutrition (NIN), Tarnaka, Hyderabad. The animals were housed in poly propylene cages with not more than 3 animals per cage, at an ambient temperature of 18 ± 20 ⁰C with 12-h-light/-dark cycle. Rats have free access to standard rodent feed (Nutrilab) and water ad libitum. The maintenance and the handling of animals were performed according to the rules and regulations of Institutional Animal Ethical Committee (Regd. No.1412 /a/11/CPCSEA).

ACUTE TOXICITY STUDIES

The ethanolic extract of Cassia auriculata L. was screened for acute toxicity following the standard method (OECD-23). Albino rats of either sex weighing about 200 – 250 g were used in this study. Animals were maintained on normal diet and water prior to and during the course of experiment. The dose of extract was prepared with 1% CMC and was administered orally. The acute toxicity was tested at the doses of 100 - 2000 mg/kg.

EVALUATION OF ANTI-DIABETIC ACTIVITY IN FLOWER OF CASSIA AURICULATA FED STREPTAZOTOCIN-NICOTINAMIDE INDUCED DIABETIC WISTAR RATS

Diabetes was induced in overnight fasted experimental groups by a single intra-peritoneal (i.p) injection of freshly prepared STZ (60 mg/kg body weight) dissolved in 0.1 M Citrate buffer (pH 4.4), 15 min after the i.p administration of NIC (120 mg/kg b.w). Hyperglycemia was confirmed by the elevated glucose levels. After 72 h, blood glucose was determined and those rats with fasting glucose levels greater than 126 mg/dl were used in the present study. Male Wistar albino rats were divided in to five groups of six animals in each group as follows: Group 1: Vehicle controlled received 1% CMC daily for 15 days. Group 2: STZ-NIC (60 mg/kg-120 mg/kg b.w) induced diabetic rats received 1% CMC daily for 15 days. Group 3: STZ-NIC (60 mg/kg-120 mg/kg b.w.) induced diabetic rats received glibenclamide (10 mg/kg, b.w., per oral). Group 4: STZ-NIC (60 mg/kg-120 mg/kg b.w,) induced diabetic rats received (200 mg/kg) Ethanolic extract of Cassia auriculata daily for 15 days. Group 5: STZ-NIC (60 mg/kg-120 mg/kg b.w) induced diabetic rats received (400 mg/kg) ethanolic extract of Cassia auriculata daily for 15 days⁸.

STATISTICAL ANALYSIS

All the values of body weights, blood glucose and biochemical estimations were expressed as mean ± SEM. The data obtained in the studies were subjected to one way analysis of variance (ANOVA) for determining the significance difference. The inter group significance was analyzed by Dunnett’s test using Graph Pad Prism Version 4. Statistical Significance i.e. difference between groups were considered at p<0.05.

RESULTS AND DISCUSSION:

In the present study the flower of Cassia auriculata L. were collected, dried and subjected to size reduction to get uniform coarse powder. The shade dried flower of Cassia auriculata successively extracted by maceration with ethanol. The percentage of yield and nature of extracts was shown in table 1. Based on the acute toxicity studies the dose of plant extract was selected for animal studies. The ethanolic extract fed streptazotocin-nicotinamide induced diabetic model shown the decreased level of blood glucose in groups IV & V. Cassia auriculata flower ethanolic extract has shown maximum reduction in blood glucose level which calculated by comparing the blood glucose level at 7^th and 14^th day of its respective groups shown in Table 2. Finally the percentage reduction of blood glucose represented ethanol extract (400 mg/kg) has shown maximum reduction in blood glucose as compared to diabetic control than other extract. Group V showed suppression of blood glucose level at 14^th day significantly (p<0.05) compared to zero day to its respective groups. The significant anti-diabetic activity of Cassia auriculata may be due to inhibition of free radicals generation and subsequent tissue damage induced by straptazotocin-nicotinamide or potentiation of insulin effect by increase either pancreatic secretion of insulin from existing beta cells.

CONCLUSION:

In the present investigation, administration of extract produced a significant reduction in blood glucose levels in Streptazotocin-Nicotinamide induced diabetic rats. More studies are required to ascertain its mechanism of action there by providing a natural hypoglycemic control treatment and thus decrease risk for diabetes.

Table: 1 Percentage of yield and physical appearance of ethanol extract:

S.No	Extract	Nature of extract	Color	Yield (% w/w)
1	ethanol	Solid	pale yellow	18.75

Table: 2 Table showing blood glucose (mg/dl) (mean ± SD) (n=6) in different groups of rats on day 0, 7&14

Days Groups	0	7	14
Vehicle control	119.7±4.27	118.2±4.11	118.5±4.43
Diabetic control	314.75± 125.25	371.5± 146.5	396.2±138.4
Glibenclamide	424.25±52.27	341.75± 124.57	304.25± 119.8
Extract I(200mg/kg)	567.5 ±43.1	483± 5.65	265±35.35
Extract II(400mg/kg)	372.3±163.2	304.6± 136.9	202.3±79

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Received on 06.07.2012 Accepted on 16.08.2012

Asian J. Pharm. Tech. 2(3): July-Sept. 2012; Page 104-106