BACKGROUND: The use of acetaminophen (APAP) is increasing recently, especially with COVID-19 outbreaks. APAP is safe at therapeutic levels, however, an overdose can cause severe liver injury. This study aims to explore possible mechanisms involved in APAP-induced hepatotoxicity and compare different hepatoprotective agents, namely vitamin E, hydrogen sulfi de (H 2 S) and necrostatin-1 (NEC-1). METHODS: Adult male albino rats were divided into groups: Control group, APAP-induced hepatotoxicity group, Vitamin E-treated group, H 2 S-treated group and NEC-1-treated group. Serum levels for aspartate aminotransferase (AST), alanine aminotransferase (ALT), interleukin-33 (IL-33), tumor necrosis factor alpha (TNF-α), reduced glutathione (GSH) and lipid profi le were measured. Histopathological examinations of liver tissue with H&E stain and immunohistochemistry for activated caspase-3 were also done. RESULTS: APAP-treated group showed elevated liver transaminases, hyperlipidemia, and defi cient liver anti-oxidative response together with disturbed hepatic architecture and increased immune-expression of activated caspase-3 in hepatic tissue. Pretreatment with vitamin E, H 2 S or NEC-1 reversed the affected parameters. Vitamin E and H 2 S showed greater improvement when compared to NEC-1. CONCLUSION: Vitamin E, H 2 S and NEC-1 showed protective effects against APAP-induced hepatotoxicity, thus they may be used as an adjuvant therapy when APAP is indicated for long periods as is the case in Fig. 2, Ref. 45).