To investigate the protective effects of kaempferol against acetaminophen in hepatocytes-induced injury. Methods:Cholecystokinin-8 (CCK-8) was used to detect the vitality of hepatocytes treated by kaempferol. The lactate dehydrogenase (LDH) released ratio was detected，which could determine the concentration and time of acetaminophen (APAP)-induced hepatocytes injury model. CCK-8 and LDH released ratio were used to detect the vitality of hepatocytes treated by APAP and kaempferol. In the meanwhile，hepatocyte morphology was to be record. P-Jun N-terminal kinase (P-JNK)，Jun N-terminal kinase (JNK)，Endo G protein expression were detected by Western Blot searching. Results:The APAP concentration was determined to be 5 mmol/L and the time was 24 hours to induce liver injury cell with drug. Compared with the APAP group，the morphology of hepatocytes was significantly improved. The ratio of apoptotic cells decreased significantly (P<0.01)，and the ratio of released LDH content decreased significantly (P<0.001). Western Blot showed that P-JNK and Endo G were significantly down-regulated. Conclusion:This experiment demonstrates that kaempferol has a protective effect on APAP-induced hepatocytes injury.