Rajiv Gandhi University of Health Sciences s160

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES

KARNATAKA, BANGALORE

M. PHARM SYNOPSIS

DECEMBER- 2011-12

“Evaluation of Anti-obesity activities of Crotalaria juncea L. in albino rats.”

BY

Mr. SREEDHAR K.S

DEPARTMENT OF PHARMACOLOGY

UNDER THE GUIDANCE OF

Dr. B.M. Vrushabendra Swamy M. Pharm, Ph. D, FICCP.

Director/Professor

Department of Pharmacology

GAUTHAM COLLEGE OF PHARMACY

SULTHANPALYA, R.T. NAGAR, BANGALORE-560032

KARNATAKA

6. BRIEF REVIEW OF THE INTENDED WORK :

6.1 INTRODUCTION:

Health is defined as soundless of physical, mental or moral condition, especially freedom from bodily pain or disease, but true health is more than that. It includes the joy of living, the power and ability to lead a satisfying and purposeful life.

Obesity and related disorders are the leading causes of illness and mortality in the developed World [1].

Effects of excess weight on morbidity and mortality have been known since the time of Hippocrates, who said,” sudden death is more common in those who are naturally fat than in the lean”[2].

The ultimate cause of obesity is an imbalance between calorie intake and energy expenditure, but the pathologic mechanisms, which lead to this imbalance are still not understood. Obesity is believed to be of both genetic and environmental origin, involving excess calorie intake, decreased physical activity, social and economic forces and metabolism and endocrine abnormalities[3].

The World Health Organization has estimated that perhaps 80% of earths 6 billion inhabitance rely upon traditional medicine for their primary health care needs, and a major part of this therapy involves the use of plant extracts or their active principles.

Crotalaria juncea L. known as Sunn or Sunn hemp, is a tropical Asian plant of the legume family (Fabaceae).Grown as a source of green manure, fodder and the lignified fiber obtained from its stem, it bears yellow flowers and elongate, alternate leaves. Sunn Hemp is also being looked at as a possible bio-fuel Annual, c. 60–250cm tall.

Many ascending branches, pubescent. Leaf simple, 2.5-10.5cm long, 6–20mm broad, linear or oblong, obtuse or subacute, apiculate, pubescent on both sides, hairs appressed, silky. Petiole 1.2-2.5mm long; stipules almost absent. Inflorescence an erect terminal and lateral raceme, up to 30cm long, 12-20-flowered. Pedicel c. 3–7mm long. Bract minute; bracteoles 2, below the calyx, 1.8-2.0cm long, pubescent, teeth linear-lanceolate. Corolla bright yellow. Vexillum ovate-oblong, slightly exserted. Fruit; 2.5-3.2cm long, sessile, pubescent, 10-15-seeded [4].

6.2 NEED FOR THE STUDY

Obesity is abnormal or excessive fat accumulation that presents risk to health. The body mass index (BMI) of a person is 25-30 kg/m2 indicates overweight and above 30 kg/m2 represents obesity. World Health Organization (WHO) assigns obesity as global epidemic. WHO’s latest study indicate that globally in 2005, approximately 1.6 billion adults (15+) were overweight and at least 400 million adults were obese. Further WHO projects that by 2015 approximately 2.3 billion people will be overweight and more than 700 million will be obese. Once it was considered that obesity was only in high income countries. But now a day, it has spread dramatically in medium and low income countries [5].

The epidemic of obesity is on the rise and this is probably due to increasingly sedentary lifestyles combined with easy availability of palatable, high fat foods. Its prevalence has shown a startling Increase in all age groups and in all the countries of the world during past 19 years. In 1980, a Department of health Survey in U.K. showed that 6% of men and 8% of women were obese, as defined by a BMI >30 kg/m2.In 1995, 15% of men and 16.5% of women were obese and more than half of the adult population is now either overweight or obese. It has been predicted that if this trend continues to the year 2005, the prevalence of obesity among men and women would by then be 18 and 24% - exactly three times the target figure. There has also been an increase in comorbid risk factors like coronary heart disease, hypertension, various cancers (e.g., endometrium, ovaries, breast, colon), gall bladder disease, NIDDM, arthritis, respiratory disease, pregnancy complications, menstrual difficulties, varicose veins, psychological problems (poor self-esteem, depression, poor employment prospects)[6,7].

The successful management of obesity is possible through lifestyle changes in diet and physical activity. But this requires extreme efforts. The market for safe and efficacious drug is therefore potentially huge, but currently available therapies are not having potential and these are associated with lot of side effects. Currently FDA approved only two drugs in US for long-term treatment for obesity. These are Orlistat and Sibutramine [8].

In severe cases, surgery is performed or an intra gastric balloon is placed to reduce stomach volume and/or bowel length, leading to earlier satiation and reduced ability to absorb nutrients from food [9].

Dietary obesity can be induced readily in laboratory rodents by giving high fat diets or cafeteria diets. Obesity also occurs in rodents given a palatable sugar solution in additional to laboratory chow. These animals consume only about half of as much chow as animals not given sugar, additional calories from sugar solution generally results in greater total dietary energy intake and development of profound obesity [10].

Recently, there has been increasing interest in the use of medicinal plants. The use of medicinal plants in modern medicine suffers from the fact that though hundreds of plants are used in the world to prevent or to cure diseases. Recently search for appropriate anti-obesity agent has been focused on plants used in traditional medicine because of leads provided by natural products that may be better treatment than currently used drugs.

There are reports that Crotalaria juncea L. is used traditionally as anti-obesity plant, so, present study selected leaves of Crotalaria juncea L. plant for evaluation of anti-obesity activity by using high fat diet induced, monosodium glutamate induced obesity and fructose induced obesity in rats.

6.3 REVIEW OF LITERATURE:

Botanical name: Crotalaria juncea L

Synonym: Crotalaria benghalensis Lam

Family: Fabaceae

Vernacular names: English: Indian hemp, Bombay hemp, Sun hemp.

Hindi: kharif, sannai sun

Tamil: sanal, sannappu

Bengali: ghore sun, shon, shonpat[11].

Telugu: Sanamachettu, Gilaka, Gilligintha[12].

Distribution Description: Native: Australia, India, although generally considered a tropical or subtropical crop, sunn hemp is drought resistant and is widely adaptable to different soil types. Its tolerance to salt and frost is low [11].

Folk uses: The seeds are said to purify the blood and are used to treat impetigo and psoriasis [11], roots are used for fever, amenorrhea, and dysentery. Leafs are used anorexia, blood disorders, amenorrhoea , obesity[12].

Fiber: The major significance of sunn hemp lies in its valuable fiber, which is extracted from the bark and used to make twine, rug yarn, cigarette and tissue papers, fishnets, sacking, canvas and cordage. Fiber is stronger when wet; it is fairly resistant to mildew, moisture and microorganisms in salt water[11].

Sunn hemp fiber has greater tensile strength and is more durable under exposure than jute. It is not as strong as hemp, sunn hemp an excellent candidate for paper-making[11].

Reported activities:

-  Antifertility activity of various extracts of Crotalaria juncea L. seeds in male mice[13].

-  Anti-inflammatory and anti-ulcerogenic effect of Crotalaria juncea L. in albino rats[14].

-  Evaluation of activity of methanolic extract of anti-diarrhoeal Crotalaria juncea L. in albino wistar rats[15].

-  Antibacterial activity of seed and flower parts of Crotalaria juncea L [16].

Chemical constituents: An Abundance of mucilage, linoleic acid (62.36%)[17], steroids, flavonoids , phenols, glycosides and triterpinoides apart from that it contains some of the interesting compounds which include monocrotaline, riddelline, seneciphylline, senecionine, trichodesmine, chodesmine, galactose specific lectin and cardiogenin 3-O-[OH]-d-xylopyranosid[18].

6.4 OBJECTIVES OF THE STUDY:

The main objective of the proposed work is to induce obesity by using high fat diet[19-21], Monosodium glutamate[22], fructose induced[23] obesity in rats and screening of Crotalaria juncea L. Leaves extract in rats.

The whole study is divided into two phase

Phase I:

-  Collection and authentication of plant material. Collected plant material subject to extraction with 70% v/v alcohol using Soxhlet apparatus with hydro alcoholic (70%v/v).

-  To investigate preliminary phytochemical constituents present in the extract.

Phase II:

To evaluate Anti-Obesity activity of Crotalaria juncea L. leaves extracts by using the experimental animal models like:

Ø  High fat diet induced obesity.

Ø  Monosodium glutamate induced obesity.

Ø  Fructose induced obesity

Parameters to be studied.

-  Body weight

-  Body temperature

-  Lipid profile(HDL, LDL, VLDL, TG, Total Cholesterol).

-  Serum glucose

-  SGOT

-  SGPT

-  Parametrial adipose tissue weight

7.0 MATERIALS AND METHODS:

7.1  Source of Data:

Whole work is planned to generate data from laboratory studies i.e. experiments are performed as described in references. Experimental studies in journals and in text books available with college and various institutions.

1.  Standard Books:

§  Goodman and Gilmann’s: The Pharmacological basis of Therapeutics.

§  H.Gerhard Vogel’s: Drug Discovery and Evaluation.

§  Katzung’s : Basic and clinical pharmacology.

§  Rang and Dale’s: Pharmacology.

§  Tortora: Human Anotomy and Physiology.

§  Guyton and Hall’s: A Text Book of Medical Physiology.

§  Wealth of India.

§  Materiamedica.

§  Glossary of Indian medicinal plants.

2.  Internet source:

§  Google

§  Pub med

§  www.sciencedirect.com

§  Wikipedia

§  SCOUPS

§  Helinet

§  Ovid

§  Open J-Gate

§  DOAJ

§  Chemical Abstracts

§  CABI

§  International Pharmaceutical Abstract

3.  Journal sources:

§  Indian Journal of Pharmacology.

§  Journal of Pharmacology and experimental Therapeutics.

§  Journal of Ethano pharmacology.

§  American Journal of Pharmacology and Toxicology.

§  Phytochemistry.

7.2ANTI-OBESITY ACTIVITY:

A.  Experimental animals: Adult albino rats weighing approximately 150-200 g will be used for anti-obesity activity. And rats are used for the acute toxicological studies. The animals will be fed with standard diet and will be given water ad libitum.

B.  Preparation of extract[15]: About 250 g of powdered shade dried leaves of Crotalaria juncea L. are subjected to successive Soxhlet extraction by using Hydro alcoholic. The prepared extracts are concentrated to lesser volume under reduced pressure and evaporate to dryness. The prepared extract is suspended in distilled water containing 2%v/v tween 80 (suspending agent).

C.  Dose: A dose of Hydro alcoholic extract will be taken as per the toxicity studies.

D.  Grouping of animals: The animals are divided into 6 groups of six rats in each group and the treatment given once.

7.3 GROUPING OF ANIMALS[25]:

In high fat diet induced obesity animals:

Experimental Animals

Albino wistar rats weighing 160-220g were divided into six groups of six in each group.

High Fat Composition:

Commercially available edible dalda (vanaspathy) and culinary grade coconut oil were obtained from local market. The high fat diet (HFD) was prepared by homogenously mixing dalda and coconut oil in the ratio of 3:2 w/w.

Induction of Obesity

Group I animals were administered with 10ml distilled water per kg body weight orally once daily for a period of four weeks by oral gavaging technique and served as negative control. For the Group II, III, IV, V, and VI in addition to normal diet and water prepared high fat diet was administered by gavaging to induce obesity. HFD was gavaged at the dose rate of 10ml per kg body weight to each animal orally daily for a period of four weeks.

Treatment Protocol

Once the Obesity was induced between 0 to 4th week of the experiment, from the beginning of fifth week to the end of the eighth week, the Hydro alcoholic extract of Crotalaria junceac L (HECJ) treatment was carried out.

Group-I: Distilled water was administered and served as negative control.

Group-II: Distilled water was administered and served as positive control

Group-III: Standard drug (Fenofibrate 200mg/kg, p.o) was administered.

Group-IV: HECJ was administered at a dose rate of 200mg/kg, p.o body

weight.

Group-V: HECJ was administered at a dose rate of 400mg/kg, p.o body

weight.

Group-VI: HECJ was administered at a dose rate of 600mg/kg, p.o body

weight.

After the completion of eighth week i.e., 56 days, on 57th day blood was collected for the estimation of biochemical parameters. Before collection of blood the animals were kept overnight fasting.

Parameters studied for this test were body temperature, bodyweights, average feed intake, weights of liver, kidneys, spleen, paramatrial adipose tissue, blood glucose, total cholesterol, HDL, LDL, VLDL, triglycerides, atherogenic index.

Parameters to be studied.

Body weight: The body weight (g) will be recorded on day 1 and then on alternate days for 40 days in each group.

Body temperature: The body temperature will be recorded on day 39 using rectal telethermo meter before and after drug administration at 30, 60, 90, 120 and 180min with a contact time of 1 minute.

Biochemical parameters: Serum glucose, SGOT, SGPT, Lipid profile (HDL, LDL, VLDL, TG, Total Cholesterol) will be measured on the 40th day in each group.

Parametrial adipose tissue weight: On the 40thday of the experiment rats will be sacrificed, parametrial adipose tissue will be dissected and weighed.

In Monosodium glutamate induced obesity:

Experimental animals:

Albino wistar rats weighing 120-150gms were divided into 6 groups of six in each group.

Induction of obesity:

Group I animals were administered with distilled water orally once daily by oral gavaging technique and served as negative control. For the Group II, III, IV, V and VI in addition to normal diet and water prepared Monosodium glutamate solution was administered by gavaging to induce obesity. Monosodium glutamate was gavaged at the dose rate of 8mg/kg body weight to each animal orally daily up to 7days.