STUDY PROTOCOL
EVALUATION OF TEST FORMULATION
FOR ANTI-ULCERATIVE ACTIVITY USING PYLORUS LIGATION INDUCED ULCER IN
EXPERIMENTAL RAT
1.0 INTRODUCTION:
Gastric ulceration is a benign lesion on the mucosal epithelium upon exposure of the stomach to excess acid and aggressive pepsin activity. It is possibly due to a disturbance in the balance between the aggressive factors like pepsin, acid, and H. pylori and the defensive factors such as bicarbonate, gastric mucus, and prostaglandins.
2.0 TEST SYSTEM DETAILS:
Species
: Rattus norvegicus
Strain : Sprague dawley; Wistar.
Age : 6-8 Weeks
Body
Wight : 150-200g
Sex : Male or Female
No.
of animals : 8 /Group
Total
animal : 56 + 5 Extra = 61
3.0 ALLOCATION OF GROUPS:
|
Groups |
Treatment |
Dose; ROA |
No. of Animals |
|
G1 |
Normal
Control |
0.9%
Normal saline /Na-CMC |
8 |
|
G2 |
Ulcerated
Control |
0.9%
Normal saline/Na-CMC |
8 |
|
G3 |
Reference
Drug- Cimetidine |
100
mpk; p.o. |
8 |
|
G4 |
Test
Formulation-1 |
X
mpk; p.o. |
8 |
|
G5 |
Test
Formulation-1 |
XX
mpk; p.o. |
8 |
|
G6 |
Test
Formulation-2 |
X
mpk; p.o. |
8 |
|
G7 |
Test
Formulation-2 |
XX
mpk; p.o. |
8 |
#
5 extra animals will be taken extra due to variability in the disease
development and possibilities of animal mortalities
4.0 METHOD:
·
Animals will be acclimatized
for 7 days prior to the main experiment and maintained all standard conditions as
per CPCSEA guidelines.
·
Animals will be randomly
divided into seven groups containing eight animals in each.
·
To induce ulcer, the fasting
rats (free access to water) will be undergoing a small surgery that is ligation
of the pylorus sphincter.
·
All animals will be
anesthetized using ketamine and xylazine (90mg/kg+10mg/kg mixture) the abdomen
will be opened by using a small incision below the xiphoid process.
·
The stomach will be exposed,
and a thread will be placed around the pyloric sphincter and tied in a tight
knot.
·
Care will be taken while tying
the knot to avoid damage involving blood vessels in the knot.
·
The rats will be put with
wide-mesh wire bottoms to prevent coprophagia during the experiment.
·
The abdomen will be sutured,
and the animals will be returned to their home cages.
·
All the drug or vehicle and
test compounds will be administered to animals 1 h before the pylorus ligation.
·
Group of animals G1 will be
served as normal control (without ulceration) and administered with 0.25%
sodium CMC at 10ml/kg.
·
Group of animals G2 will be
served as ulcerated control and administered with 0.25% sodium CMC at 10ml/kg.
·
Group of animals G3 will be
served as a reference drug and administered with cimetidine at the dose of
100mg/kg p.o.
·
Similarly, groups of animals G4
to G7 will be administered with test compound/test sample at different dose
levels.
·
After 4h of pylorus ligation
all animals will be euthanized by overdosing with an anesthetic agent
(halothane).
·
The stomachs will be removed,
and the contents will be drained into tubes and centrifuged at 1000 rpm for 10
minutes. The supernatant will be then subjected to analysis for gastric volume,
pH, and acidity
·
Cleaned stomachs will be
scored using a dissecting microscope with a square-grid eyepiece based on grading
on a 0–5 scale (depicting the severity of vascular congestions and
lesions/hemorrhagic erosions).
·
Mean ulcer score for each
animal will be expressed as ulcer index (U.I) and the percentage of inhibition
against ulceration will be determined using the expressions:
U.I.
= [Ulcerated area/total stomach area] × 100.
%Ulcer
inhibition = [U.I. in control − U.I. in test] × 100/U.I. in control
5.0 End Point Parameter:
· Ulcer Index
· % Ulcer inhibition
Free Acidity and Total Acidity Calculation
· Histopathology of stomach tissue
· pH and acidity of GI content
· Superoxide dismutase (SOD) and malondialdehyde (MDA) (In stomach homogenate)
6.0 REFERENCE(S):
6.1 Alefe
Norahun Mekonnen, Seyfe Asrade Atnafie, and Mohammedbirhan A. Wahab Atta.
Evaluation of Antiulcer Activity of 80% Methanol Extract and Solvent Fractions
of the Root of Croton macrostachyus Hocsht: Ex Del. (Euphorbiaceae) in Rodents.
Evidence-Based Complementary and Alternative Medicine Volume 2020, Article ID
2809270, 11 pages https://doi.org/10.1155/2020/2809270.
6.2 Vivek
Srivastava, A.H.M.
Viswanathaswamy, and Govind Mohan.
Determination of the antiulcer properties of sodium cromoglycate in
pylorus-ligated albino rats. Indian J
Pharmacol. 2010 Jun; 42(3): 185–188.
6.3 Woretaw
Sisay Zewdu, Tezera Jemere Aragaw. Evaluation of the Anti-Ulcer Activity of
Hydromethanolic Crude Extract and Solvent Fractions of the Root of Rumex
nepalensis in Rats. Journal of Experimental Pharmacology 2020:12
Das SK, Pansuriya PV, Shukla ST, Gohil KJ, Roy SP, Choudhury A, Sutariya VN. Preclinical evaluation of Vallaris solanacea (roth) kuntze stem for its antiulcer and antioxidant activity in wistar albino rats. Oriental Pharmacy and Experimental Medicine. 2014 Mar;14(1):7-13. DOI:10.1007/s13596-013-0133-3
END OF DOCUMENT
4
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment