EVALUATION OF HEPATOPROTECTIVE ACTIVITY OF HERBAL FORMULATION(S) USING ANTI TUBERCLOSIS DRUGS INDUCED HEPATOTOXICITY IN EXPERIMENTAL ANIMALS

                                                                  STUDY PROTOCOL

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

1.0 INTRODUCTION

Tuberculosis (TB) is one of the major communicable diseases. Isoniazid (INH) and rifampicin (RIF) are the two major regimens currently used for the treatment of TB for a period of 4 to 6 months, which may induce hepatotoxicity. The incidence of hepatic dysfunction is more, when INH and RIF are used in combination. The liver dysfunction is due to the synergistic effect of INH and RIF. Hydrazine (HYZ) a metabolite of INH is converted to toxic compound by CYP450, which leads to hepatotoxicity. RIF, aggravates hepatotoxicity by inducing CYP450, as a result more toxic metabolites are generated from hydrazine. 

2.0 TEST SYSTEM DETAILS:

Species : Rattus norvegicus (Rats)

Strain : Wistar or Sprague Dawley

Age : 8-10 weeks

Body Wight : 120-200 g

Sex : Male or Female

No. of animals : 8 /Group

3.0  ALLOCATION OF GROUPS:

Groups	Treatment	Dose; ROA	No. of Animals
G1	Normal Control	0.25% Na- CMC; qdX8W	8
G2	Disease Control	0.25% Na- CMC; qdX8W	8
G3	Positive Control- Silymarin	100 mpk; qdX8W	8
G4	Plant Extract/Herbal Formulation-1	X mpk, p.o	8
G5	Plant Extract/Herbal Formulation-1	XX mpk, p.o	8
G6	Plant Extract/Herbal Formulation-2	X mpk, p.o	8

2 extra animals will be taken extra in study due to variability in the disease development and possibilities of animal mortalities  

4.0 METHOD:

·  Rats weighing 150-200gm will be included in the present investigation.

· The animals will be housed in polypropylene well-aerated cages at normal atmospheric temperature (21 ± 3°C) and normal 12 h light/dark cycle.

· Rats will be free access to water and supplied daily with laboratory standard diet of known composition ad libitum.

· Animals of group G1 and G2 will be administered with 0.25% sodium carboxymethylcellulose (CMC).

· All the animals (except G1) will be administered with Isoniazid (31 mg/kg), Rifampicin (62 mg/kg b.wt) and Pyrazinamide (155 mg/kg) for 7 weeks to induce chronic liver and kidney injury.

· Animals of Group G3 will be received reference drug Silymarin at the dose of 100mg/kg b.wt p.o.

· Animals of group G4, G5 and G6 will be administered with test sample at different dose level.

· All the animals will be treated for 7 weeks with vehicle or Silymarin or test samples at different dose levels.

· Animals will be anaesthetized and blood will be collected on week 2, 3, 4, 5, 6 and 7th for Albumin, Globulin, Total Bilirubin, AST, ALT, Total protein, albumin, ALP, LDH analysis.

· Further, at the end of the experiment all the animals will be humanely sacrificed, liver and kidneys will be isolated weighed and stored in 10 % buffered formalin and further processed for histopathology.

5.0 END POINT PARAMETER(S):

·   Clinical observation

· Feed water consumption

· Body Weight

· Liver Function Profile - Albumin, Globulin, Total Bilirubin, AST, ALT, Total protein, albumin, ALP, LDH, GGT and CRP

· Anti-oxidant Profile – Catalase (CAT), malonaldehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione (GSH).

· Liver Lysosomal enzymes- Cathepsin D and β-Galactosidase

· Histopathology of Live tissue

6.0  REFERENCE(S):

6.1 Ahmed Wahid, Ashraf N. Hamed, Heba M. Eltahir and Mekky M. Abouzied. Hepatoprotective activity of ethanolic extract of Salix subserrata against CCl4- induced chronic hepatotoxicity in rats. BMC Complementary and Alternative Medicine (2016) 16:263

6.2 M. Sankar, Johanna Rajkumar, and Dorai Sridhar Hepatoprotective Activity of Heptoplus on Isoniazid and Rifampicin Induced Liver Damage in Rats. Indian Journal of Pharmaceutical Sciences. Received 2014 Mar 20; Revised 2015 Jan 31; Accepted 2015 Sep 14.

6.3 Chao Wang, Rui-Qin Fan, Yan-Xiang Zhang, Hao Nie, Kan Li. Naringenin protects against isoniazid- and rifampicin induced apoptosis in hepatic injury. World J Gastroenterol 2016 November 28; 22(44): 9775-9783.

                                       END OF DOCUMENT

You may like to read theses links:   

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

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

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

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