Abstract: This study aimed to investigate the ameliorative effects of 60% ethanol elution fraction (ESMW) from Si Miao Wan on the hepatic lipid accumulation and its mechanism.TG kit, BODIPY fluorescence staining, QPCR, WB, oil red O staining, and AMPKα knockdown were used to detect the ability of ESMW to improve lipid accumulation in hepatocytes stimulated with free fatty acid.Furthermore, the effects of ESMW on the oral glucose tolerance, serum biochemical indexes, TG content in liver tissue, the expressions of mRNA and protein related to lipid metabolism in liver tissue were studied in mice fed with high fat diet to verify the mechanism of ESMW fraction on hepatic lipid accumulation.The results showed that ESMW inhibited lipid accumulation induced by free fatty acids by regulating AMPK signaling pathway, and that ESMW significantly improved the lipid metabolism of mice fed with high fat diet, with relation to AMPK signaling pathway.