AIM:The potential of Trifolium pratense(red clover) extract in the prevention of lipid disorder has attracted increasing attention in recent years.In this study,the aim was to determine whether and how red clover extract affected the development of murine diet-induced non-alcoholic steatohepatitis.METHOD:Non-alcoholic steatohepatitis was induced in C57BL/6 mice by feeding mice with a methionine-choline-deficient(MCD) diet.Hematoxylin and eosin staining was used for histological analyses.Real-time PCR was used to analyze the mRNA expression levels.RESULTS:Hepatic steatosis and necroinflammation was observed in MCD diet-fed mice,and this diet-induced steatosis was significantly attenuated,whereas liver inflammation was not significantly attenuated,by red clover extract treatment.Consistent with the results of H&E staining,the MCD diet-induced increase of liver triglycerides and cholesterol levels were significantly reduced by red clover extract treatment.However,with the improvement in hepatic steatosis,mRNA levels of acetyl CoA oxidase,carnitine palmitoyl transferase-1,and liver fatty acid-binding protein,three genes regulated by peroxisome proliferator-activated receptor(PPAR) α,were unaffected.CONCLUSION:Red clover extract alleviated MCD diet-induced hepatic steatosis,but did not ameliorate liver inflammation in C57BL/6 mice,and the improvement in hepatic steatosis was not through activating PPARα. AIM: The potential of Trifolium pratense (red clover) extract in the prevention of lipid disorder has sought increased in recent years. In this study, the aim was to determine whether and how red clover extract affected the development of murine diet-induced non -alcoholic steatohepatitis. METHOD: Non-alcoholic steatohepatitis was induced in C57BL / 6 mice by feeding mice with a methionine-choline-deficient (MCD) diet. Hematoxylin and eosin staining was used for histological analyzes. Real-time PCR was used to analyze the mRNA expression levels .RESULTS: Hepatic steatosis and necroinflammation was observed in MCD diet-fed mice, and this diet-induced steatosis was significantly attenuated, whereas liver inflammation was not significantly attenuated, by red clover extract treatment. Consistent with the results of H & E staining, the MCD diet-induced increase of liver triglycerides and cholesterol levels were significantly reduced by red clover extract treatment. However, with the improvement in h epatic steatosis, mRNA levels of acetyl CoA oxidase, carnitine palmitoyl transferase-1, and liver fatty acid-binding protein, three genes regulated by peroxisome proliferator-activated receptor (PPAR) alpha, were unaffected. CONCLUSION: Red clover extract alleviated MCD diet- induced hepatic steatosis, but did not ameliorate liver inflammation in C57BL / 6 mice, and the improvement in hepatic steatosis was not through activating PPARα.