Aim: Hepatotoxicity is the most serious adverse effect of rifampicin (RIF). We aimed to investigate the potential hepatoprotective effect of mannose-functionalized poly(lactic-co-glycolic acid)(PLGA)/RIF nanoparticles (NPs) in rats as a possible promising approach to minimize RIF-induced hepatotoxicity. Ma- terials & methods: Mannose-functionalized PLGA/RIF NPs were fabricated and characterized in vitro, then the hepatoprotective effect of optimized NPs was studied on rat and cell culture models. Results: Follow- ing intraperitoneal administration of RIF NPs into rats, highly significant differences in levels of serum transaminases and oxidative stress markers, associated with significant differences in expression of Bax and Bcl-2 genes between NP- and free RIF-treated groups, revealing the hepatoprotective potential of NPs. Conclusion: RIF NPs may represent a promising therapeutic approach for tuberculosis via reducing dose frequency and consequently, RIF-induced hepatotoxicity.