PROTOCOL
EVALUATION OF MUCOPROTECTIVE
ACTIVITY OF SHARK
SQUALENE AGAINST ETHENOL-INDUCED MUCOSAL DAMAGE IN RATS
1.0 OBJECTIVE OF STUDIES :
Identification, Characterization of Shark squalene.
To investigate mucoprotective activity of Shark squalene on Ethanol-induced mucosal damage models in rats.
2.0 TEST SYSTEM DETAILS:
Species : Rattus norvegicus (Rat)
Strain : Wistar or Sprague Dawley
Age : 8-10 weeks
Body Wight :
120-200 g
Sex : Male/Female
No. of animals : 8
/Group
Total Animals : Model (40+05 Extra= 45)
3.0 ALLOCATION OF GROUPS:
|
Groups |
Treatment |
Dose;
ROA |
No. of
Animals |
|
G1 |
Normal Control |
Normal saline or 0.25% Na-CMC |
8 |
|
G2 |
Negative Control |
Pyloric ligation group |
8 |
|
G3 |
Reference Drug- Ranitidine |
30 mg/kg; p.o. |
8 |
|
G4 |
Test Formulation-1 |
X mg/kg; p.o. |
8 |
|
G5 |
Test Formulation -2 |
XX mg/kg; p.o. |
8 |
*The
doses and ROA (Routes of administration) will be decided based on the type of reference
drug
4.0 METHOD:
In this method, the rats will be treated for 7 days with Shark squalene.
After seven days of treatment, animals fasted
for 24 hours with free access to water.
Shark squalene and Standard drugs will
be given orally before the administration of Ethanol.
The animals will be then killed 4
hours after the operation under the overdosage of anesthetic ether and
gastric contents are collected.
After centrifugation, the gastric
contents of each animal are individually assessed for gastric acidity, acid
volume, pH, and mucin content.
The inhibition of gastric mucosa is quantified.
Lesions score will be defined as the
sum of the length (mm) of gastric lesions observed and measurement of the length of the
lesions will be performed.
5.0 END POINT PARAMETER(S):
·
Clinical observation
·
Feed water consumption
·
Body Weight.
·
The stomach will be
isolated, weighed
·
The contents are subjected
to centrifugation [3000rpm for 10min] and analyzed for volume, pH, free and total acidity, ulcer score ulcer
index, gastric wall mucus content mucin content.
·
Antioxidant-activity-
Lipid peroxidation & Catalase of Gastric tissue.
·
Measuring the protein, gastric
mucosal barrier glutathione, lipid peroxidase, alkaline phosphate, etc.
·
Histopathological investigations
of gastric mucosa in mucosal-damaged rats.
6.0 REFERENCE(S):
6.0 Shark squalene (shark liver oil) from VitaDigest. [online]. 2007 [cited
2009 Oct 9]; Available from: URL: http://www.vitadigest.com/vitamins--supplements-squalene.html
7.0 Solomon N, Passwater R, Joelssun I, Haimes L. Shark liver oil; Nature’s
Amazing Healer. Kensington books; 1997.
8.0 Shark oil monograph from, Natural
medicines comprehensive data base. Scientific Gold
Standard for Evidence-Based clinical information on natural medicines. Therapeutic research faculty, Stockton: CA
95208; Nov 20, 2009.
9.0 Pugliese PT, Jordan K, Cederberg H, Brohult J. Some biological
actions of alkoxyglycerols from Shark liver oil. J Altern
Complement Med.1988; 4:8799.
10.0
Lewkowicz N, Lewkowicz
P,Kurnatowska A, Tchorzewski H. Biological actions and Clinical applications of
Shark liver oil. Pol Merkur Lekarskj 2006;20(119):598-601.
11.0
Riber C , Wojdemann M, Bisgaard
T, Ingels H, Rehfeld JF, Oslen O. Fish oil reduces gastric acid secretion. Scand J
Gastroenterol 1999;34:845-8.
12.0
Mycek MJ, Harvey RA, Champe PC.
Lippincott’s Illustrated Review, Pharmacology. 2nd ed. Philadelphia (NY): Lippincott-Raven Publishers; 1997. p. 236-41.
13.0
Nutrasanus. [online]. 2009
Available from: URL: http://www.nutrasanus.com/shark-liver-oil.html
14.0
Shark squalene (shark liver
oil) from VitaDigest. [online]. 2007 [cited 2009 Oct 9]; Available from: URL: http://www.vitadigest.com/vitamins--supplements-squalene.html
15.0
Solomon N, Passwater R, Joelssun
I, Haimes L. Shark liver oil; Nature’s Amazing Healer. Kensington books; 1997.
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