关键词: 非酒精性脂肪性肝病, HepG2, 丹酚酸B, 棕榈酸, 自噬, 体外

Abstract: Objective To induce the steatosis of HepG2 cells by palmitic acid （PA） in vitro to establish a nonalcoholic fatty liver disease （NAFLD cell model,and to observe the effect of salvianolic acid B （Sal B） and autophagy inhibitor of 3-methyladenine（3-MA） intervention on intracellular steatosis. Methods HepG2 cells were divided into control,model （PA intervention）,intervention （PA plus Sal B intervention） and inhibitor （3-MA, PA and Sal B intervention） groups. The best intervention concentrations of PA and Sal B were screened by MTT method,red O staining was used to observe lipid droplets in each group of cells and the fluorescence intensity of autophagy marker,LC3B was detected by fluorescence microscopy. Western blot was used to detect the expression of LC3B protein. Results The TC,TG,ALT and AST levels in supernatant of the model group were（0.57±0.07） mmol/L,（0.99±0.07） mmol/L,（98.47±7.00） IU/L and （88.36±8.54） IU/L,significantly higher than [（0.14±0.02） mmol/L,（0.26±0.03） mmol/L,（23.37±2.24） IU/L and（27.27±3.19） IU/L,respectively,P<0.01],which were also higher than [（0.30±0.04） mmol/L,（0.56±0.06） mmol/L,（53.36±5.33） IU/L and （56.37±7.66） IU/L in intervention group or （0.43±0.02） mmol/L,（0.83±0.10） mmol/L,（86.84±3.37） IU/L and （75.82±3.43） IU/L in the inhibitor group,P<0.05];the absorbance value of oil red O in the model group was （0.666±0.009）,significantly higher than （0.247±0.011） in the control group,or （0.477±0.013） in intervention group or （0.507±0.002） in the inhibitor-intervened group （P<0.001）;the expression of LC3B protein in intervention group was（0.97±0.01）,significantly stronger than 【（0.22±0.02）,P<0.01】 in the model group,while it decreased a little,e.g.（0.44±0.05） in inhibitor-intervened group. Conclusion Sal B might reduce the lipid accumulation in HepG2 cells by increasing intracellular autophagy.

Key words: Nonalcoholic fatty liver disease, HepG2, Salvianolic acid B, Palmitic acid, Autophagy, In vitro