EVALUATION OF HEPATOPROTECTIVE OF TEST FORMULATION AGAINST ANTI-TUBERCULAR DRUGS INDUCED HEPATOTOXICITY

 

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EVALUATION OF HEPATOPROTECTIVE OF

TEST FORMULATION AGAINST ANTI-TUBERCULAR  DRUGS  INDUCED  HEPATOTOXICITY

 

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 (8+5 = 40)

 

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 (Isoniazid-7.5 mg/kg, Rifampicine-10 mg/kg, and Pyrazinamide-35 mg/kg)	0.25% Na-CMC	8
G3	Reference Drug- Sylimarin	100 mg/kg; p.o.	8
G4	Test Formulation-1	X1 mg/kg; p.o.	8
G6	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

 

3.0  METHOD:

Model: Antitubercular drugs induced hepatic damage in rats.

      (a)  Antitubercular drugs induced peroxidation damage in liver:

·       The animals will be divided into control, negative control, positive control and and two test formulation groups (different doses).

·       Hepatotoxicity will be induced by administration of three anti-tubercular drugs (Isoniazid-7.5 mg/kg, Rifampicine-10 mg/kg, and Pyrazinamide-35 mg/kg) for 35 days by intra gastric route in sterile saline.

·       After the experimental period, blood will be collected from the inner canthus of the eye from each animal under isofuran aneasthesia using caplillary tube, clear serum will be separated at 2500 rpm for 10 min.

·       Serum will be subjected to standard biochemical estimations for different parameters like total bilirubin (TB) , Direct bilurubin, Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Alkaline phosphates (ALP)  and total protein (TP).

(b)  In-vivo hepatoprotective activity evaluation :

·       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).

·        A portion of liver tissue in each group will be fixed in 10% formasal (formalin diluted to 10% with normal saline) and will be studied for histopathological changes.  

     

4.0  END POINT PARAMETER(S):

·       Clinical observation

·       Feed water consumption

·       Body Weight.

·       Liver marker enzymes :

AST, ALT, ALP, Total Bilirubin, Direct Bilirubin, Total proteins,

·       Liver antioxidant enzymes.

a)     Enzymatic antioxidant

                                                                                   I.     Catalase

                                                                                 II.     Superoxide dismutase

b)     Non-enzymatic antioxidant

                                            I. Reduced Glutathione

5.0   REFERENCE(S):

6.0  Wolf PL. Biochemical diagnosis of liver diseases. Indian J Clinical Biochem 1999;14:59–90.

7.0  Hasegawa R.,Chujo T, Sai-Kato K, Umemura T, Tanimura A, Kurokawa Y. Preventive effects of green tea against liver oxidative DNA damage and hepatotoxicity in rats treated with 2-nitropropane. Food. Chem. Toxicol. 1995;33: 961-70.

8.0   Hussain ZA,Amresh G, Satyawan SB, Chandana VR. A hepatoprotective activity of Amaranthus spinosus in experimental animals. Food  Chem Toxicol 2008; 46:3417–21.

9.0  Shukla ST, Kulkarni VH, Habbu PV, Jagadeesh KS, Patil BS, Smita DM. Hepatoprotective and antioxidant activities of crude fractions of endophytic fungi of Ocimum sanctum Linn. in rats. Oriental Pharmacy and Experimental Medicine. 2012 Jun;12(2):81-91.

10.0                   Sheweita SA, Abd El-Gabar M, Bastawy M. Carbon tetra chloride changes the activity of cytochrome P450 system in the liver of male rats: role of antioxidants. Toxicology. 2001; 169: 83-92.

11.0                   Aruoma OI. Nutrition and health aspects of free radicals and antioxidants. Food Chem Toxicol 1994;32:671–83.

12.0                   Rao GMM, Rao, CV,  Pushpangadan  P,  Shirwaikar A. Hepatoprotective effects of rubiadin, a major constituent of Rubia cordifolia Linn. J Ethnopharmacol  2006; 103:484–90.

13.0                   Agarwal RR, Dutt S. Chemical examination of cuscuta reflexa Rosb Part I. The constituents. J Indian Chem Soc 1935;1:384-88.

14.0                   Petri WA. Goodman and Gilmans.The pharmacological Basis of Therapeutics.10^th edition.New York:McGraw-Hill;2001:1273-312.

 

 

 

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