This study was carried to evaluate the protective effect of rutin and N-acetyl cysteine (NAC) against isoniazid-induced (INH) hepatotoxicity. In this study, Seventy two adult male rats were divided into six equal groups as follow: control group: 1 ml saline /Kg b.wt I/P. INH group: 100 mg INH /kg b.wt I/P. INH + Rutin group: 100 mg INH /kg b.wt I/P with 200 mg rutin /kg orally. INH + NAC group: 100 mg INH /kg b.wt I/P with 300 mg NAC /kg orally. Rutin group: 200 mg rutin /kg/day orally. NAC group: 300 mg NAC /kg b.wt orally. Serum and tissue specimens were collected at 7th, 14th and 21th day. IP injection of Isoniazid induced significant increase in values of ALT, AST, ALP, GGT and total bilirubin, while significant decrease in total protein and albumin was recorded. INH + Rutin showed significant decreases in the levels of ALT, AST, ALP and GGT with significant increase in albumin values and significant increase in total protein however significant decrease in the level of total bilirubin was recorded only at 21 days. INH + NAC induced significant decreases in the levels of ALT, AST, ALP and GGT with significant increase in albumin and total protein at 14 and 21 days and significant decrease in total bilirubin at 21 days. Rutin and NAC have potent protective effect against INH induced hepatotoxicity.