“Anti-obese activity of Butea monosperma bark extract in experimentally induced obese rat ”.
M. Pharmacy Dissertation Protocol Submitted to
Rajiv Gandhi University of Health Sciences, Karnataka
Bangalore – 560 041
By
Mr. Patel Dixit B. Pharm.
Under the Guidance of
Mr.T.PRAKASH M.Pharm
Asst. Professor
Department of Pharmacology
P.G. Department of Pharmacology,
Acharya & B.M.Reddy College of Pharmacy,
Soladevanahalli, Chikkabanavara
Hesaraghatta Main Road, Bangalore – 560 090
RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE, KARNATAKA
ANNEXURE-II
PROFORMA FOR REGISTRATION OF SUBJECT FOR DISSERTATION
1 / Name of the candidateand address
/ Mr. DIXIT KUMAR
S/O Sri Dilipbhai patel
A/P –Ubharan,
Tal- Malpur
Dist- Sabarkantha,
State- Gujarat, India
Pin-383335
2 / Name of the institution / Acharya & B.M. Reddy College Of Pharmacy.
Soladevanahalli, Hesaraghatta Road, Chikkabanavara Post,
Bangalore -560090.
Phone No: 080 65650815
3 / Course of study and subject / M. Pharm.
Pharmacology.
4 / Date of admission / June-2007.
5 / Title of the topic / “Anti-obese activity of Butea monosperma bark extract in experimentally induced obese rat ”.
6 / Brief resume of intended work
6.1 Need of the work
6.2 Review of Literature
6.3 Objective of the study / Enclosure I
Enclosure II
Enclosure III
7 / Materials & Methods
7.1 Source of data
7.2 Methods of collection of data ( Including sampling ,procedure if any)
7.3 Does the study require investigation on animals?
If yes give details
7.4 Has ethical clearance been obtained from your institution in case of 7.3 / Enclosure IV
Enclosure V
Enclosure VI
Yes (Copy Enclosed)
8 / List of references / Enclosure VII
9 / Signature of the candidate
10 / Remarks of the guide
11 / Name & Designation of
11.1 Guide
11.2 Signature of Guide
11.3 Head of the Department
11.4 Signature of HOD / Mr. T.PRAKASH M.Pharm
Asst. Professor
P.G. Department of Pharmacology,
Acharya & B.M.Reddy College of Pharmacy,
Soladevanahalli, Chikkabanavara
Hesaraghatta Main Road , Bangalore – 560 090
Prof. Kalyani Divakar M.Pharm. (Ph.D)
P.G. Department of Pharmacology,
Acharya & B.M.Reddy College of Pharmacy,
Soladevanahalli, Chikkabanavara
Hesaraghatta Main Road , Bangalore – 560 090
12 / Remarks of the Principal
12.1 Signature / PRINCIPAL
Dr. Divakar Goli
Acharya & B. M. Reddy College of Pharmacy,
Soladevanahalli, Hesaraghatta main road,
Bangalore-90.
Enclosure - I
6. BRIEF RESUME OF INTENDED WORK
6.1 NEED OF THE WORK:
Obesity is a worldwide epidemic characterized by excess adipose tissue that contributes to numerous chronic diseases and early mortality.1,2 Obesity affects an estimated 300 million people worldwide and has a profound, negative impact on health and quality of life among adults, adolescents, and even children in the United States and other industrialized countries. The attention that this epidemic receives both nationally and internationally is justifiable based upon obesity’s detrimental impact on health, the enormous economic burden it imposes, and its increasing prevalence. In the United States, it is estimated that approximately 64.5% of adults can be classified as overweight or obese.3
Abdominal obesity signifies excess adipose tissue located in the abdomen, and is believed to contribute disproportionately to ill health. The adverse health consequences associated with abdominal obesity, as well as obesity in general, are vast,4 and include cardiovascular disease,5,6 stroke, type 2 diabetes mellitus,7 hypertension, dyslipidemia, liver diseases, gallbladder disease, osteoarthritis,10,11 respiratory problems such as asthma12 and sleep apnea,13 as well as cancers of the breast, endometrium, prostate, and colon.8,9 Furthermore, aerobic capacity and the ability to perform physical activities may be hindered by obesity,14,15 and activities of daily living may be difficult to perform. In addition to this increased morbidity, approximately 325,000 deaths in the United States each year among nonsmokers are attributable to obesity.16
The most commonly used method today for classifying an individual as overweight or obese is based upon body mass index (BMI), a value that is determined by dividing body weight (in kilograms) by the square of height (in meters). The World Health Organization defines overweight among adults as a BMI 25.0 kg/m2, and obesity as a BMI 30.0 kg/m2, regardless of sex. The use of BMI has gained international acceptance because of the associations between BMI and adiposity,17 BMI and disease risk,4 and BMI and mortality.18 The major limitation of BMI, however, is that it does not differentiate between weight that is fat and weight that comprises lean body mass. Furthermore, BMI does not provide information about fat distribution, and therefore does not identify abdominal obesity. Due to these limitations, an older adult who has experienced the common loss of muscle tissue with aging may have a BMI value in the healthy range despite muscle wasting and excess abdominal fat. In addition, some athletes are misclassified as overweight using BMI because of the relative density of lean body mass. These limitations necessitate the use of another measure to estimate health risk associated with increased body fat.
The survey of literatures reveals that the parts of Butea monosperma (Linn) i.e. bark is bitter, astringent, cooling, laxative, useful in the ulcer, jaundice, elephantiasis, obesity, urinary disorders, fever, epilepsy. 19
An extensive literature survey from all scientific sources revealed that no work has been done on the anti-obese effect of the bark of the Butea monosperma. Hence the present study is planned to investigate the anti-obese activity of the bark of Butea monosperma.
Enclosure – II
6.2 REVIEW OF LITERATURE:
The name of the plant is Butea monosperma (Linn) belongs to the family of Fabaceae. In different places in India it is called as “Palash”(Bengali), “Muttuga”(Kannada),“Palaasu” (Tamil),“Palasamu”(Telugu),“Khakharo”(Gujarati). 19
Description of plant:
It is a middle sized, deciduous tree growing up to 12-15 m height and 1.3-2 m in girth. The trunk is somewhat crooked, has light brown or bluish grey bark, and exudes a ruby- red gum when incised. Leaves are alternate, long–petiole, 3- foliolate, and large, 25-45 cm long. Flowers are large, orange red or scarlet on racemes 15-20 cm long on bare branches. Pods are silvery white, 10-15 cm long, pendulous containing one large, flat kidney shaped seed. Mature seed are glossy, wrinkled, and deep reddish brown in color. Plants flower from January to April depending on the locality.19
Distribution of the plant:
Plant is common throughout the India extending on the north-west Himalayas and common all over Bengal, southern India, Myanmar and Sri lanka.30
Chemical constituents:
The bark contains several pyrone and furan derivatives, and related compound like flavons, chalkones, aurones, and their glycoside. It should be made of butein and its glucoside butrin and palasonin because of their biological activity. A few unusual compounds such as allophonic acid derivative and lactone have been reported.
It also contains fatty acid and several lipolytic and proteolytic enzyme. Steroid stigmasterol and its glucoside and triterpene 3-hydroxyeuph 25-ene have been also isolated and contain carbohydrate base, leucocyanidin, gallic acid, B vitamin, riboflavin, thiamine and cyanidin.19
It aiso contain fixed oil 18%, albuminoid substances 19%, and glucose 6%. Fatty acid isolated from this oil is oleic, linoleic, linolenic, palmitic, steric, arachidic, behenic and lignoceric acid.29
Traditional Therapeutic Uses 19:
Ø Its roots are used in elephantiasis and in eye defect.
Ø Its flowers are used in urinary calculi, as anti-septic and jaundice, inflammation.
Ø Its seed is used in urinary disorders, skin disease and in ulcer.
Ø Its bark used to treat intestinal worm infection, obesity, internal tumor and persistent dysentery.
Ø Mothers for any backache use its gum after childbirth and in epilepsy.
Scientific Work:
Ø Anti-diabetic potential of Butea monosperma in rats.20
Ø Anti-fungal constituent from the stem bark of Butea monosperma.21
Ø Anti-convulsive activity of Butea monosperma flowers in laboratory animals.22
Ø Efficacy of Butea monosperma on dermal wound healing in rats.23
Ø Anthelmintic activity of Butea monosperma.24
Ø Anti-diarrhoeal activity of Butea monosperma in Experimental Animals.25
Ø Anti-inflammatory activity of Butea monosperma flowers.26
Enclosure – III
6.3 OBJECTIVE OF THE STUDY:
Ø Extraction with different solvents (Petroleum-Ether, Chloroform, Ethanol).
Ø Preliminary Phytochemical analysis of different extract.
Ø Toxicity study and calculation of LD50.
Ø Evaluating the efficiency of Butea monosperma extract in anti-obese activity.
Enclosure – IV
7. Materials and Methods
7.1 SOURCE OF DATA:
The source of data will be obtained for experiments which involves.
Ø Laboratory based studies
Ø Literature survey, CD ROM, Chemical abstracts
Ø National & International Journal
Ø Textbooks.
Ø Internet.
Enclosure – V
7.2 METHOD OF COLLECTION OF DATA
Field and laboratory studies
Fieldwork:
The bark of the selected plant (Butea monosperma.) will be collected from forest of Bangalore rural district.
Laboratory work:
Animals
Either sex of Wistar albino Rats (150-200 gm) will be selected for Anti-obese activity. The animals will be housed in standard environmental condition & provided with food & water ad libitum.
Solvents selected for extraction:
Petroleum ether.
Chloroform.
Ethanol.
Preliminary & Phytochemical analysis:
Different extracts of the plant are analyzed for preliminary tests and Phyto-chemical constituents.31
Data analysis
Statistical analysis of the data will be evaluated by analysis of variance (ANOVA) followed by Dunnets test.
Methodology:
(1) Extraction:
The bark of Butea monosperma will be collected, shade dried. Then they are coarsely powdered & extracted by petroleum ether, chloroform and ethanol to get sufficient crude extract. Extract is subjected to concentrate. Then alcoholic extracts are subjected for pharmacological evaluation of Anti-obese activity.
(2) Toxicity study:
Determination of LD50 carried as per OECD guidelines 425 using animal.
(3) Pharmacological study design.
Ø Evaluation of Anti-obese activity:
1. Cafeteria diet induce obesity:28
Cafeteria diet (CD) consist of 3 diets (condensed milk, 48g + bread, 48g), (chocolate, 18g + biscuits, 36g + dried coconut, 36g) and (cheese, 48g + boiled potato,60g).
Ø Group I: Normal control.
Ø Group II: Cafeteria diet control, which receive CD along with normal pellet chow and water ad libitum.
Ø Group III: CD + Low dose of EBM.
Ø Group VI: CD + Medium dose of EBM.
Ø Group V: CD + High dose of EBM.
Ø Group VI: CD + Sibutramine (5 mg/kg, p.o./ day).
The diet is given for 40 days at the and following parameters are determined.
Ø Body Weight.
Ø Food Intake.
Ø Serum glucose and lipid levels.
Ø Body temperature.
Ø Locomotor activity.
Ø Organ and fat pad weight.
The result will be compared with those obtained in normal control group.
Table-1 Cafeteria diet induces obesity in Wistar albino rats.
Groups / Treatment / Dose / No of Animals in group / Animals Sacrificed or NotI / Control Vehicle / 5 ml/kg / 6 / Sacrificing
II / Cafeteria Diet Control / 25g/day/rat / 6 / Sacrificing
III / Cafeteria Diet + Ethanolic extract / Low dose / 6 / Sacrificing
IV / Cafeteria Diet + Ethanolic extract / Medium Dose / 6 / Sacrificing
V / Cafeteria Diet + Ethanolic extract / High dose / 6 / Sacrificing
VI / Cafeteria Diet + Standard drug.(Sibutramine) / 5 mg/kg, p.o./ day. / 6 / Sacrificing
Total No of Animals : 36
Sacrificing Animals : 36
2. Atherogenic diet induce obesity:28
Atherogenic diet (AD) consists of Cholesterol:1%, Cholic acid: 0.5% and Lard oil: 5 %
Ø Group A: Normal control.
Ø Group B: Atherogenic diet control which receive AD along with normal pellet chow and water ad libitum.
Ø Group C: AD + Low dose of EBM
Ø Group D: AD + Medium dose of EBM
Ø Group E: AD + High dose of EBM
Ø Group F: AD + Sibutramine (5 mg/kg, p.o./ day)
The diet is given for 40 days at the end following parameters will be determined.
Ø Body Weight.
Ø Food Intake.
Ø Serum glucose and lipid levels.
Ø Body temperature.
Ø Locomotor activity.
Ø Organ and fat pad weight.
The result will be compared with those obtained in normal control group.
Table-2 Atherogenic diet induces obesity in Wistar albino rats.
Groups / Treatment / Dose / No of Animals in group / Animals Sacrificed or NotA / Control Vehicle / 5 ml/kg / 6 / Sacrificing
B / Atherogenic Diet Control / 25g/day/rat / 6 / Sacrificing
C / Atherogenic Diet + Ethanolic extract / Low dose / 6 / Sacrificing
D / Atherogenic Diet + Ethanolic extract / Medium Dose / 6 / Sacrificing
E / Atherogenic Diet + Ethanolic extract / High dose / 6 / Sacrificing
F / Atherogenic Diet + Standard drug.(Sibutramine) / 5 mg/kg, p .o./ day. / 6 / Sacrificing
Total No of Animals: 36
Sacrificing Animals: 36
3. Monosodium –L-glutamate (MSG) induce obesity 27:
In this young albino rat between the ages of 2 to 40 days are used and treated with single doses of 0.1- 6.0mg/kg body weight of monosodium-L- gluconate for five consecutive days. Control rat are treated with physiological saline. Then animal are weaned at 3 week of age, housed under control temperature and artificial light/dark cycle and provided with commercial powder chow and tap water at libitum.
Ø Group1: Normal control.
Ø Group 2: Monosodium -L- gluconate (MSG) control.
Ø Group 3: MSG + Low dose of EBM.
Ø Group 4: MSG + Medium dose of EBM.
Ø Group 5: MSG + High dose of EBM.
Ø Group 6: MSG + Sibutramine (5 mg/kg, p .o./ day).
Then following parameter will be determined:
Ø Body Weight.
Ø Food Intake.
Ø Serum glucose and lipid levels.
Ø Body temperature.
Ø Locomotor activity.
Ø Organ and fat pad weight.
The result will be compared with those obtained in normal control group.
Table-3 Monosodium –L- glutamate induce obesity in Wistar albino rats.
Groups / Treatment / Dose / No of Animals in group / Animals Sacrificed or Not1 / Control Vehicle / 5 ml/kg / 6 / Sacrificing
2 / Monosidium-L- glutamate induce control / 0.1- 6.0 mg /kg body weight / 6 / Sacrificing
3 / Monosidium-L- glutamate+ Ethanolic extract / Low dose / 6 / Sacrificing
4 / Monosidium-L- glutamate+ Ethanolic extract / Medium Dose / 6 / Sacrificing
5 / Monosidium-L- glutamate+ Ethanolic extract / High dose / 6 / Sacrificing
6 / Monosidium-L- glutamate + Standard drug. (Sibutramine) / 5 mg/kg, p .o./day. / 6 / Sacrificing
Total No of Animals: 36
Sacrificing Animals: 36
Ø The data obtained from the above study will be subjected to statistical analysis using ‘t’ test.