The study was designed to evaluate the protective effect of wheat germ oil (WGO) against hepatotoxicity induced during treatment with the combination of two anti-tubercular (anti-TB) drugs, Isoniazid (INH) and Rifampicin (RIF) in rats. Fatty acid and Vit E. contents of WGO were assayed by GC-MS and HPLC respectively. Rats received WGO at two different doses (250 and 500 mg/kg. body weight, p. o, for 30 days) 30 minutes before Anti-TB drugs. Liver functions test, inflammatory mediator marker (IL-10 and NF-κB), oxidative stress (GSH, MDA) and NO were determined with different techniques. Furthermore, histological findings, immunohistochemical and ultra-structure were carried out. In-vitro analysis of the WGO revealed that linoleic and oleic acids were the major compounds, and WGO was rich in vitamin E. Significant elevation in liver enzymes, NF-κB, and depletion in IL-10, along with a disturbance in the antioxidant defense systems. Meanwhile, WGO improves all these changes in a dose-dependent manner as compared with the anti-TB intoxicated group. The hepatoprotective effect of WGO was confirmed with immune histochemical and histopathological findings. WGO has a hepatoprotective effect against hepatotoxicity induced by combined therapy of anti-TB drugs (INH+RIF).