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: -
- 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.
- 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.
- Obesity will be induced in
overnight fasted rats by High Fat Diet.
- 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.
- At the end of the study, all
animals will be sacrificed with 150 mg/kg of Thiopentone sodium 24 h after
the final treatment.
- 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 (0th,30th,60
days)
2. Blood cholesterol (0th,30th,60 days)
3. Blood serum lipase (0th,30th,60 days)
4. Serum
adiponectin (0th,30th,60
days)
5. Serum
SGOT, SGPT, ALP (0th,30th,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
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