Genistein plays an active role in improving nonalcoholic fatty liver disease (NAFLD). This study is designed to investigate the effect of genistein on liver inflammation in rats with nonalcoholic steatohepatitis (NASH). Forty SPF male SD rats were randomly divided into normal group, model group, genistein low-dose group (0.1% wt/wt) and high-dose group (0.2% wt/wt) with 10 rats in each group. After 12 weeks’ feeding, liver tissues and serum samples of rats were taken, and HE staining was used to perform pathological examination of liver tissues, then the degree of inflammatory infiltration was observed and NAFLD activity score(NAS) was calculated. With corresponding kits, several indicators were detected, namely, serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), liver TC and TG, and serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood glucose and serum endotoxin. The levels of tumor necrosis factor (TNFα) in liver and insulin in blood of rats were detected by enzyme linked immunosorbent assay (ELISA), then the HOMA-IR index was calculated. Immunohistochemistry staining was used to observe the expression level of TLR4 protein and the RT-PCR was used to detect Tlr4 mRNA expression in liver tissue. The results showed that genistein could reduce TLR4 protein and gene expression, decrease the endotoxin and TNFα, alleviate the inflammatory reaction and make the indicators detected in blood and liver stay near normal in NASH rats. In conclusion, genistein can ameliorate hepatic inflammatory reaction in nonalcoholic steatohepatitis rats.