Objectives The purpose of the study was to prepare an effective and new drug delivery system for enhancing the stability of Phenethyl isothiocyanate (PEITC), and its hepatoprotective effect in the carbon tetrachloride (CCl4)-induced damage in hepatocellular carcinoma G2 (HepG2) cell line via nuclear factor (erythroid-derived 2)-like 2 (Nrf2) pathway. Methods Gold nanoparticles were synthesized and then characterized by XRD, SEM, SEM-EDX analysis, hydrodynamic diameter and zeta potential measurements. 1.0024 mM PEITC, a naturally occurring isothiocyanate, an active ingredient was loaded onto the characterized AuNPs. The cytotoxicity test of PEITC-AuNP and effects on ALT, AST, Nrf2 levels and total antioxidant capacity (TAC) of CCI4-induced HepG2 cells were investigated. Results PEITC-AuNPs and PEITC decreased ALT and AST levels (p0.05). PEITC didn’t increase the Nrf2 level in CCI4-induced HepG2 cells. TAC of both PEITC-AuNPs and PEITC administration increased significantly compared with CCl4 group (p Conclusions PEITC-AuNPs were more effective than PEITC which resulted in more hepatoprotective and antioxidant effects via Nrf2 activation against CCl4-induced liver injury in HepG2 cells.