Salvianolic acid B (Sal B) the most abundant bioactive member in Salvia miltiorrhiza and has been reported lots of benefits on the treatment of cardio-cerebral vascular diseases and metabolic diseases. Lipid droplets are dynamic organelles, excessive lipid droplets accumulation in liver caused fatty liver disease. In this study, we are interested in the effect of Sal B on hepatic lipid accumulation and its possible mechanism. We found Sal B treatment significantly decreased lipid accumulation and TG level in primary hepatocytes, this is the first demonstration for the effect of Sal B on lipid droplets. Meanwhile, a stimulation of Sal B on autophagy of hepatocytes was discovered, further we used autophagy inhibitor 3-MA and reveled that after administration of 3-MA Sal B showed little effect on improving hepatic lipid accumulation, this means the effects of Sal B on lipid drop depend on autophagy. This study demonstrates Sal B ameliorate hepatic lipid accumulation through activation of autophagy. These findings contribute to its benefit on liver disease related to hepatic lipid accumulation and hepatic autophagy.