EVALUATION OF HEPATOPROTECTIVE AND ANTIOXIDANT ACTIVITY OF TEST FORMULATION IN CCL4 –INDUCED HEPATIC DAMAGE IN RATS

STUDY PROTOCOL

 

EVALUATION OF HEPATOPROTECTIVE AND ANTIOXIDANT ACTIVITY OF

   TEST FORMULATION IN CCL₄ –INDUCED HEPATIC DAMAGE IN RATS’’ 

1.0   TEST SYSTEM DETAILS:

Species                  : Rattus norvegicus (Rat)

Strain                    : Wistar or Sprague Dawley

Age                       : 8-10 weeks

Body Wight          : 150-200 g

Sex                        : Male/Female

No. of animals      : 8 /Group

Total Animals       : Model (40+10 Extra= 50)

 

2.0   ALLOCATION OF GROUPS:

Model :

Groups	Treatment	Dose; ROA	No. of Animals
G1	Normal Control	Normal saline or 0.25% Na-CMC	8
G2	Negative Control (CCL4 i.p. dose)	0.25% Na-CMC	8
G3	Reference Drug- Silymarin	100 mg/kg; p.o.	8
G4	Test Formulation-1	X1 mg/kg; p.o.	8
G5	Test Formulation -1	XX1 mg/kg; p.o.	8

 

*The doses and ROA (Routes of administration) will be decided based on the type of reference drug

# 10 extra (~20%) animals will be taken extra due to CCL₄ Induction possibilities of animal mortalities

 

3.0  METHOD:

Model: Carbon tetra chloride induced hepatic damage in rats.

      (a) CCl₄ induced peroxidation damage in liver:

·       Albino Wistar rats weighing 150-200 g, will be maintained in standard environmental condition and fed with standard laboratory diet and water ad libitum will be used for the experiment.

·       CCl₄ will used to induce hepatotoxicity in rats.

·       Different Test formulations will be administered orally at different dosage level (for specified period of time) to study the efficacy against CCl₄ induced peroxidative damage in liver of rats.

      (b)  In-vivo hepatoprotective activity evaluation :   

v  Animals will be sacrificed by cervical dislocation (only in codition of body weight in < 200 g above 200 g use Thiopental sodium 150 mg/kg), livers excised and the following parameters will be estimated. 

v  Serum glutamate pyruvate transaminase (SGOT)

v  Serum glutamate oxaloacetate transaminase (SGPT)

v  Serum Alkaline phosphatase (SALP)

v Total Bilirubin

v Total Protein

v Total Triglycerides

v Total cholesterol

 

4.0  END POINT PARAMETER(S):

·       Clinical observation

·       Feed water consumption

·       Body Weight.

·       Histopathological studies : The pancreas will be collected from the animals and fixed in 10% formalin, and subjected to histopathological examination.

·       Endogenous enzymatic and non-enzymatic antioxidant levels : In-vitro free radical scavenging activity : 2,2-diphenyl-1-picrylhydrazyl [DPPH] Scavenging activity Hydroxyl radical scavenging activity

·      In-vivo free radical scavenging activity :   Animals will be sacrificed by cervical dislocation, livers excised and the following parameters will be estimated Glutathione, Total thiol, Lipid peroxidation, Catalase

 

 

5.0   REFERENCE(S):

1.0  Razvi SU. Effect of Ocimum sanctum Linn. leaf extract on hepatotoxicity induced by antitubercular drugs in rats. Indian J Physiol Pharmacol 2003; 47(04):465-70.

2.0  Guo B, Wang Y, Sun X, Tang K. Bioactive natural products from endophytes: A Review. Appl Biochem Microbiol 2008;44(2):136-42.

3.0  Weber LW, Boll M, Stampfl A. Hepatotoxicity and mechanism of action of haloalkanes: carbon tetrachloride as a toxicological model. Crit Rev Toxicol 2003;33(2):105-36.

4.0  Achliya GS, Koyagale NR, Wadodkar GS, Dorle AK. Hepatoprotective activity of Panchagavya Ghrita against carbon tetrachloride induced hepatotoxicity in rats. Indian J Pharmacol 2003;35:308-11.

5.0       ShuklaST, Kulkarni VH, Habbu PV, Jagadeesh KS, Patil BS, Smita DM. Hepatoprotectiveand antioxidant activities of crude fractions of endophytic fungi of Ocimumsanctum Linn. in rats. Oriental Pharmacy and Experimental Medicine. 2012Jun;12(2):81-91.

6.0   Hukkeri VI, Akki KS, Sureban RR, Gopalakrishna B, Byahatti VV, Rajendra SV. Hepatoprotective activity of the leaves of Nyctanthes arbor-tristis linn. Indian J Pharm Sci 2006;68:542-3

7.0                   Prabhakar KR, Veerapur VP, Parihar Vipan K, Priyadarsini KI. Evaluation and optimization of radioprotective activity of Coronopus didymus Linn. In g-Irradiated Mice. Int J Radiat 2006;82(8):525-36.

8.0                   Ghosh MN. Fundamentals of Experimental Pharmacology. 2^nd ed. Culcutta:  Scientific Book Agency; 1984. p.154-7.

                                         END OF DOCUMENT

You may like to read theses links:   

1. List of All SOPs and Documents for In-vivo Laboratory

2. List of All SOPs and Documents for Study Protocol

3. List of Pharmacological Study Models with details about disease Inducing agent, Standard or Reference Drug and related parameters.

4.List of OECD Guidelines or Toxicological Studies

5.List of Pharmacopeial Test or Quality Control Test or Batch Testing Parameters (In-vivo)

6.EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF HERBAL FORMULATION(S) USING ALCOHOL INDUCED HEPATOTOXICITY IN EXPERIMENTAL ANIMALS

7.EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF HERBAL FORMULATION(S) USING ANTI TUBERCLOSIS DRUG RIFAMPICIN, ISONIAZIDE AND PYRAZINAMIDE INDUCED HEPATOTOXICITY IN EXPERIMENTAL ANIMALS

 

SHARE

Owner

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.

• 
• 
• 
• 
•