1.0 TEST SYSTEM DETAILS:
Species : Rattus norvegicus (Rats)
Strain : Sprague Dawley or Wistar
Age : 6-7 weeks
Sex : Female
No. of animals: 8 /Group
Total animals: 56
2.0 TEST ARTICLES DETAILS
HB1: Herbal Formulation-1
3.0 VEHICLE DETAILS
The test articles will be formulated by utilizing 0.5 % Methyl Cellulose as the vehicle.
4.0 ALLOCATION OF GROUPS
|
Group No. |
Group Description |
Disease Induction procedure |
Treatment administered |
Dose Volume and Route |
|
G1 |
Normal Control |
The animal will be subcutaneously
injected with distilled water for 10 consecutive days. |
0.5% Na-CMC, p.o., b.i.d. |
5 ml/kg, p.o. |
|
G2 |
Disease Control |
Subcutaneous injection of Estradiol
benzoate (EB)-5 mg/kg on the first day, 3 mg/kg on the second day to the ninth
day, and then 5 mg/kg on the tenth day. Oxytocin 3U/kg will be administered
by i.p.2 hours after the last administration of EB. |
0.5% Na-CMC, p.o., b.i.d. |
|
|
G3 |
Reference Control |
Ibuprofen -100 mg/kg, p.o., b.i.d. |
||
|
G4 |
Treated with a low dose of HB-1 |
HB-1:
5-15 mg/kg, b.i.d. |
||
|
G5 |
Treated with intermediate dose 1 of
HB-1 |
HB-1: 15-50 mg/kg, b.i.d. |
||
|
G6 |
Treated with intermediate dose 2 of
HB-1 |
HB-1: 50-150 mg/kg, b.i.d. |
||
|
G7 |
Treated with a high dose of HB-1 |
HB-1: 150-500 mg/kg, b.i.d. |
5.0 METHOD:
· Healthy animals will be selected for the study, randomized based on body weight, and will be assigned to 7 groups consisting of 8 animals each.
· Normal Control (animals assigned to group G1) will receive 0.5% Na-CMC, p.o., b.i.d.
· Animals of group G2 will serve as Disease Control-receiving vehicles used for formulating HB1
· Animals of group G3 will be treated with reference Standard- Ibuprofen -100 mg/kg, p.o., b.i.d.
· Animals of group G4-G7 will be treated with HB1, at different incremental concentrations as outlined in Section 4.0 of Annexure-I, twice daily, orally.
· All groups except G1 will receive subcutaneous EB injections (5 mg/kg on day 1, 3 mg/kg on days 2-9, and 5 mg/kg on day 10) to facilitate induce disease, followed by i.p. administration of oxytocin 3U/kg 2 hours later on the tenth day, and writhing behavior will be recorded within 30 minutes of oxytocin injection.
· After the writhing response test, the rats will be euthanized and their uterine tissues will be harvested for histopathology. Some parts of the tissues will be quickly stripped, homogenized, and stored at -80°C for further analysis.
6.0 PARAMETERS TO BE EVALUATED:
· Body weight: Twice a week.
· Determination the of Levels of NO, Ca++, Estradiol, Prostaglandin E2 and, F2-a (PGF2α and PGE2).
· PGF2α to PGE2 ratio
· iNOS activity in rat uterine tissues
· Histopathological studies.
2.0 REFERENCE(S):
1. Bi, W., Zhou, J., Zhao, L., Wang, C., Wu, W., Zhang, L., … Zhou, F. (2021). Preventive effect of different citrus essential oils on primary dysmenorrhea: in vivo and in vitro study. Food Bioscience, 42, 101135. doi:10.1016/j.fbio.2021.101135
2. Yang, Lu, Zhengyu Cao, Boyang Yu, and Chengzhi Chai. "An in vivo mouse model of primary dysmenorrhea." Experimental Animals 64, no. 3 (2015): 295-303.
No comments:
Post a Comment