Evaluation of Anti-Obesity Activity of Test Formulation Using High Fat Diet-induced Obesity in Experimental Animal

                                                                          Study Protocol

Evaluation of Anti-Obesity Activity of Test Formulation Using High Fat Diet-induced Obesity in Experimental Animal 

Test System

Animal                                   : Albino Rat,

Strain                                      : Wistar

Sex                                          : Female

Age                                         : Adult Rat

Number of animals                 : 35 (Rat)

No. of animals /group             : 7 per group (Rat)

No. of groups                          : 6 (Rat)

Acclimatization                      : 7 days

Identification of animals        : Marking the animals with picric acid

Animal husbandry

Room Temperature                : 23±2 °C

Relative humidity                   : 50-70 %

Light/dark cycle                     : 12-hourly

Feed                                       : Feed purchased from a commercial supplier will be provided ad libitum to the animals.

Water                                      : Filtered drinking water will be provided ad libitum.

 

 

Experimental Design:

(I) Normal Control: This group will be treated as a control group.

(II) Disease Control: In this group, Wistar albino rats will be given High Fat Diet.

.(III) Positive Control: In this group, Simvastatin will be given 10 mg/kg/day, p.o. for 60 days after inducing obesity with a High Fat Diet.

(IV) Treatment Group I: In this group Test Formulation will be given in 200 mg/kg, p.o. for 60 days after inducing obesity with High Fat Diet on the first day of the study

(V) Treatment Group II: In this group Test Formulation will be given in 600 mg/kg, p.o. for 60 days after inducing obesity with a High Fat Diet on the first day of the study 

(VI) Formulation Group III- In this group Test Formulation will be given in 1500 mg/kg, p.o. for 60 days after inducing obesity with a High Fat Diet on the first day of the study

Group No.	Group Details	Disease Induction & Regimen	Drug Dose	No. of Animals
G1	Normal Control	NA	0.9 % Normal saline	6
G2	Disease Control	High Fat Diet	100 mg/kg	6
G3	Positive Control	High Fat Diet+ Simvastatin	100 mg/kg +10 mg/kg	6
G4	TF-1	High Fat Diet	100 mg/kg +200 mg/kg	6
G5	TF-2	High Fat Diet	100 mg/kg + 600 mg/kg	6
G6	TF-3	High Fat Diet	100mg/kg + 1500mg/kg	6

Procedure: -

1. The body weight of individual animals will be taken daily for each group and records will be maintained from the starting day of the study till the last dosing will do and also before sacrificing the animal.
2. If the death of any animal occurs in between the study time, its weight will also be taken. Food intake will be measured each day.
3. Obesity will be induced in overnight fasted rats by High Fat Diet.
4. The development of obesity will be confirmed by triglycerides, cholesterol level, serum lipase, serum adiponectin, serum SGOT, SGPT, and ALP level on the weekly basis.
5. At the end of the study, all animals will be sacrificed with 150 mg/kg of Thiopentone sodium 24 h after the final treatment.
6. The liver and adipose tissue will be removed intact, washed with saline, and blotted dry on filter paper. The liver will then be cut into two equal pieces. One half will be stored at −80°C to measure liver MDA and GSH; the remaining part of the liver and whole adipose tissue will be fixed for 24 h in 10% buffered formaldehyde for histopathological examination.

 

ENDPOINT PARAMETERS

1.     Blood triglycerides level (0^th,30^th,60 days)

2.     Blood cholesterol (0^th,30^th,60 days)

3.     Blood serum lipase (0^th,30^th,60 days)

4.     Serum adiponectin (0^th,30^th,60  days)

5.     Serum SGOT, SGPT, ALP (0^th,30^th,60 days)

6.     Antioxidant estimation in the liver (MDA, GSH, SOD, CATALASE)

7.     Leptin

8.     Histopathological examination of liver and adipose tissue

 

 References:

1.  C. Roh and U. Jung, “Screening of crude plant extract with anti-obesity activity,” International Journal of Molecular Sciences, vol.13, no. 2, pp. 1710–1719, 2012.

2. B. M. Spiegelman and J. S. Flier, “Obesity and the regulation of energy balance,” Cell, vol. 104, no. 4, pp. 531–543, 2001.

3. P. G. Kopelman, “Obesity as a medical problem,” Nature, vol. 404, no. 6778, pp. 635–643, 2000.

4. S. Panico and A. Iannuzzi, “Dietary fat composition and the metabolic syndrome,” European Journal of Lipid Science and Technology, vol. 106, no. 1, pp. 61–67, 2004.

5. D. Nath, M.-T. Heemels, and L. Anson, “Obesity and diabetes,”Nature, vol. 444, no. 7121, p. 839, 2006.

6. A.M. Gajda, “High-fat diets for diet-induced obesity models. Open diet purified formula for rats,” Obesity, 9 pages, 2009.

7. M. J. Heikens, A. M. Gorbach, H. S. Eden, et al., “Core body temperature in obesity,” American Journal of Clinical Nutrition, vol. 93, no. 5, pp. 963–967, 2011

                                                         

                                                   END OF THE DOCUMENT

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