INTRODUCTION:

World health organization (WHO) has recommended the evaluation of traditional plant treatment for diabetes as they are effective, non- toxic, with less or no side effects and are best for oral therapy¹. Mostly people prefer herbal medicine rather than synthetic ones therefore there is a need for the search for an effective and safe drug for the treatment of diabetes. Bombax ceiba is commonly known as silk cotton tree and semal which belongs to family Bombacaceae. The tree has a straight tall trunk and its leaves are deciduous in winter. The tree bears red flower with 5 petals appearing in the spring before the new foliage². Dried leaf extracts of the plant were subjected to chemical investigation, which led to the isolate one known compound mangiferin^3,4.

Number of traditional uses has been reported in the Indian traditional systems of medicine such as Ayurveda, Siddha and Unani. Flower and bark of B. ceiba has been reported for hypotensive and Hypoglyceamic⁵, analgesic⁶, antioxidant⁷, Anti-inflamatory⁸, hepatoprotective⁹, Immunomodulatory¹⁰, antimicrobial¹¹ activities. Also it was used for the treatment of sexual debility, bleeding woundsand vaginal infections¹².Since there is no scientific evidence found for Hypoglycemic potential of B. ceiba leaves extract where as leaves are frequently used in tribals of Aravali region, so in the present study, an attempt has been made for evaluation of antioxidant and Hypoglycemic potential of different extracts of B. ceiba leaves.

MATERIAL AND METHODS:

Procurement of raw material and its authentication

Plant Material: The plant Bombax ceiba was collected from upper ki Oden, Aravali mountain region in Nathdwara during the month of April to June 2013.The leaves were identified by Dr. K.P. Dahariwal, Botanist at S.M.B Govt. PG. College Nathdwara. The leaves were taken and dried in shade. Then the shade-dried leaves were made into coarse granules and were used for different investigation.

Preparation of different Extracts

The shade dried leaves of Bombax ceiba were powdered and defatted with petroleum ether at 60° to 65°C. Extraction was performed by using Ethyl acetate, n-Butanol and Hydroalcoholic (30:70) solvents by Soxhaltion process. Extracts were dried under vacuum pressure and kept in desicatior for further study.

Phytochemical screening:

Phytochemical examinations¹³ were carried out for all the extracts as per the standard methods to find out the presence of various phytoconstituents and depicted in Table 2.

Toxicological Studies

Albino rats of Wistar strain (150-200 gm body weight) of either sex and healthy young adult female Swiss albino mice, nulliparous, non-pregnant and weighing 25-32 gm procured from in house animal facility of Institute. All animals were housed under standard conditions of temperature 25±2 0C, 65±10% relative humidity, 12:12 hr Light: Dark cycle and fed with standard pellet diet (Trimurti foods, Udaipur, India) and water ad libitum. Experimental protocols were approved by Institutional animal ethical (Registration No. 918/05/CPCSEA) of B. R. Nahata College of Pharmacy-SIRO, Mandsour-458001 (M.P.) India.

Determination of Acute toxicity (ALD₅₀)

The acute toxicity was determined in albino mice, maintained under standard conditions. The animals were fasted overnight prior to the experiment. Fixed dose (OCED Guideline No. 420) method of CPCSEA was adopted for toxicity studies¹⁴.

Three mice in each group were randomly selected. They were treated with aqueous suspension of Extracts i.e. Ethyl acetate extract (ETE), Butanolic Extract (BTE) and Hydroalcoholic Extract (HAE) at 2,000 mg/kg body weight as a single oral dose. The animals were observed for morbidity and mortality at 1, 2, 4, and 6 hr on the day of dosing and once a day thereafter up to 14 days. The study was performed in accordance with Organization for Economic Co-operation and Development (OECD) test guideline No.420.

Table 1: Acute toxicity study of different extracts of Bombax ceiba

Treatments	Dose (mg/kg)	No. of animals used	Mortality	% Mortality
Ethyl acetate extract of Leaves of Bombax ceiba (ETE)	2000	3	0	0
Butanolic extract of Leaves of Bombax ceiba (BTE)	2000	3	0	0
Hydroalcoholic Extract of Leaves of Bombax ceiba (HAE)	2000	3	0	0

Hypoglycemic Activity

Groups:

Group 1 (Control): Normal Saline

Group 2 (Standard): Glibenclamide

Group 3 (ETE400): Ethyl Acetate extract of Bombax ceiba extract 400mg/kg bw

Group 4 (BTE400): n-Butanolic extract of Bombax ceiba extract 400mg/kg bw

Group 5 (HAE400): Hydroalcoholic extract of Bombax ceiba extract 400mg/kg bw

Hypoglycemic activity of Bombax ceiba on normoglycemic rats

Normoglycemic Study¹⁵ were carried out in 12 hrs fasted normal rats, which were equally divided into five groups of five rats each. The normal control group received only vehicle (Normal Saline solution) and standard group received 1 ml of reference drug (Glibenclamide 2 mg/kg bw.), while group from third to fifth were administered with 1 ml of Extracts (ETE, BTE and HAE, at 400 mg/kg) Blood samples were collected from tail vein prior to dosing (0 Hr.) and then at regular intervals of 1 hr, 2 hr and 3 hrs respectively and subjected to fasting blood glucose level.

Study on Oral Glucose Tolerance Test (OGTT)

OGTT¹⁶ of plant extracts was carried out in overnight fasted normal rats, which were equally divided into five groups of five rats each. Group of normal control received only vehicle (1 ml of Normal Saline) and standard group received 1 ml of reference drug Glibenclamide (2mg/kg), while group from third to fifth were administered with 1 ml of Extracts (ETE, BTE and HAE at dose 400 mg/kg). Thereafter, following 30 min post extract administration all the animals were fed with glucose (2 g/kg). Blood samples were collected from tail vein prior to dosing and then at 30, 60, 90 and 120 min after glucose administration. The fasting blood glucose level was analyzed using glucose-oxidase-peroxide reactive strips (Accu-chek, Roche Diabnostics, GmbH, Germany).

Statistic Analysis

The values were represented as mean ± SD. and the data obtained from this study was subjected to one-way analysis of variance (ANOVA) followed students “t” test.

RESULT AND DISCUSSION:

Result reviled that Hydroalcoholic Extract shown better yield 10.4% w/w compare to other followed by Ethyl acetate 10.1% w/w, and n-Butanol9.9 % w/w. Phytochemical Screening suggest that Alkaloids, saponins, and sterols are absent in all three extracts and Sugar, Flavonoids, Phenolic compound, and Tannins present in all three extracts. For Toxicological evaluation it is found that all extracts (ETE, BTE and HAE) showed no mortality on 2000mg/kg. Therefore 2000mg/kg dose was considered as a safe dose, so 1/10^th and 1/5^th(200mg and 400mg) of that were we can select for all in vivo experiments as sub maximal and maximal dose. Here maximal dose used for the study. Result for Hypoglycemic activity on Normoglycemic rats’ model suggest that Blood glucose level significantly reduced by HAE and shown has better potential as compare to other extracts and Standard (Glibenclamide). (Table 2 and Fig.1).

Study of different extract on Oral Glucose Tolerance Test (OGTT) suggest the significant reduction in Blood glucose level in rats and found that HAE was superior to other extracts (Table 3 and Fig. 2), and shown 25.40% reduction in Blood glucose level

Table 2: Blood Glucose level Reduction in Normoglycemic Rats Model

Treatment	Blood glucose Level(mg/dl)				% Reduction
Treatment	0 Hour	1Hour	2 Hours	3 Hours	% Reduction
Control	69.48± 0.137	70.2± 0.125	70.4±0.175	70.1±0.015	---------
Standard	70.96±0.313	59.08±0.042	53.48±0.087	43.94±0.258	37.32
ETE( 400mg/kg)	71.46±0.288	66.16±0.183*	61.18±0.417*	58.42±0.117*	16.69
BTE(400 mg/kg)	70.58±0.107	68.48±0.162*	63.5±0.185*	55.86±0.243*	20.31
HAE(400mg/kg)	69.86±0.083	61.22±0.642*	57.32±0.312*	53.18±0.187*	24.13

Mean ± SEM for n=5, *p<0.05

Table 4: Blood Glucose level Reduction at OGTT Rats Model

Treatment	Blood glucose Level(mg/dl)					% Reduction
Treatment	0 Min.	30 Min. (After glucose Administration)	60 Min. (After glucose Administration)	90 Min. (After glucose Administration)	120 Min. (After glucose Administration)	% Reduction
Control	69.48± 0.17	117.8± 0.125	111.4±0.175	101.1±0.015	91.4±0.187	---------
Standard	70.96±0.161	102.08±0.141	97.48±0.087	71.94±0.258	60.1±0.117*	34.24
ETE (400mg/kg)	71.46±0.288	129.16±0.163*	109.18±0.117*	91.42±0.227*	71.01±0.231*	22.30
BTE (400mg/kg)	70.58±0.107	121.48±0.192*	119.5±0.185*	92.86±0.423*	72.11±0.271*	21.11
HAE (400mg/kg)	69.86±0.083	132.22±0.441*	121.32±0.112*	95.18±0.287*	68.18±0.287*	25.40

Mean ± SEM for n=5, *p<0.05

CONCLUSION:

Oxidative stress and lipid peroxidation are early events related to radicals generated during the renal impairment. Also the generation of reactive oxygen species has been proposed as a mechanism by which many chemicals can induce nephrotoxicity increases the lipid peroxidation and suppresses the antioxidant defense mechanisms in renal tissue. Now bombax has proved that anti diabetic potential on Normal rats possibly due to regenerate β cells. Study on Chemical constituents and their mechanism related to the Diabetic care has to be established. Also diabetic associated problems like Nephrotoxicity, Cardiotoxicity and Retinotoxicity study of different extracts and their mechanism has to be required to Study. Further investigation of individual compounds and characterization for the observed significant efficacy also needed

ACKNOWLEDGEMENT

Authors are Thankful to Dept. of Science and Technology (DST) Jaipur for their financial support to this research.

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Received on 27.01.2016 Modified on 14.02.2016

Research J. Pharm. and Tech. 9(3): Mar., 2016; Page 205-208

DOI: 10.5958/0974-360X.2016.00036.6