Objective To investigate the role and mechanism of angiopoietin⁃like protein 8（ANGPTL8） in N⁃diethylnitrosamine（DEN）⁃induced acute liver injury in mice. Methods Male wild⁃type（WT）and ANGPTL8 knockout（ANGPTL8 KO）C57BL/6J mice were selected and injected with DEN（50 mg/kg）intraperitoneally to induce acute liver injury models（32 mice in each group at 15 days of age and 14 mice in each group at 8 weeks of age）. The expression of ANGPTL8 in liver tissue and primary liver cells were detected by PCR and immunofluores⁃ cence；the levels of ALT and AST in serum were detected by the kit；the histopathological changes of liver were observed by HE staining；in the accumulation of reactive oxygen species（ROS）in primary liver cells was examined by flow cytometry and confocal microscopy；the levels of inflammatory factors IL⁃6 and IL⁃1β serum were detected by ELISA；the apoptosis of hepatocytes were detected by TUNEL staining. Results The expression of ANGPTL8 in the liver was significantly up⁃regulated at 3 days after DEN stimulation. Compared with WT mice，ANGPTL8 KO could significantly down⁃regulate the levels of ALT，AST，IL⁃6 and IL⁃1β in the serum induced by DEN；the hepatic lobular disorder，inflammation⁃related cell infiltration，hepatocyte congestion，vacuolar degeneration and necrosis induced by DEN were significantly improved in ANGPTL8 KO mice；the ROS accumulation and hepatocytes apoptosis induced by DEN were dramatically induced in ANGPTL8 KO mice. Conclusion DEN up⁃regulates the expression of ANGPTL8 in the liver，and ANGPTL8 further promotes the accumulation of ROS，the expression and secretion of inflammatory factors and hepatocyte apoptosis，which eventually aggravates DEN⁃induced acute liver injury.