摘要
Salvianolic acid B(Sal B)is a polyphenolic antioxidant that has been shown to have anti-lipid accumulation,anti-inflammatory,and free oxygen radical scavenging activities in various diseases.Here,we aimed to examine whether Sal B could alleviate non-alcoholic fatty liver disease(NAFLD)and explore the possible mechanisms.Signaling pathways involved in oxidative stress,including SIRT3,SOD2,and FOXO1 pathways,were investigated by Western blotting analysis,RT-qPCR,and immunoprecipitation(IP).In the present study,oleic acid(OA)successfully induced lipid and peroxide accumulation,decreased SIRT3 expression,and increased FOXO1 acetylation.However,Sal B significantly reversed these trends.SIRT3 plasmid transfection further reduced the expression of acetylated FOXO1 and considerably enhanced the regulation of SIRT3 and acetylated FOXO1 induced by Sal B.Following SIRT3 siRNA transfection,Sal B-induced down-regulation of acetylated FOXO1 was blocked,suggesting that Sal B-mediated protection occurred through SIRT3-mediated FOXO1 deacetylation.The SIRT3/FOXO1 pathway was a critical therapeutic target for controlling oxidative stress in NAFLD,and Sal B conferred protection against OA-induced hepatic steatosis and oxidative stress through SIRT3-mediated FOXO1 deacetylation.
丹酚酸B(Salvianolic acid B,Sal B)是一种多酚类抗氧化剂,已被证明在多种疾病中具有抗脂质积累、抗炎和清除氧自由基的活性。我们旨在研究Sal B是否可以改善非酒精性脂肪性肝病(Non-alcoholic fatty liver disease,NAFLD)的疾病进展并探索其可能的机制。通过油酸诱导HepG2细胞建立NAFLD模型组细胞,Sal B干预模型细胞建立治疗组细胞,进行生化分析及油红O染色检测各组细胞内脂质含量,通过免疫荧光及流式细胞仪检测细胞内活性氧(reactive oxygen species,ROS)含量;试剂盒及酶标仪定量细胞内脂氧化物丙二醛(Malonydialdehyde,MDA)的含量;通过蛋白质印迹分析、RT-qPCR和免疫沉淀(Immunoprecipitation,IP)检测参与氧化应激的信号通路蛋白,包括SIRT3、SOD2和FOXO1通路蛋白。研究发现,油酸(Oleic acid,OA)可以诱导细胞内脂质、过氧化物及脂氧化物的积累,降低SIRT3的表达,并促进FOXO1乙酰化。然而Sal B治疗后显著扭转了这些趋势。进一步通过转染SIRT3质粒及SIRT3 siRNA发现,SIRT3质粒转染后降低了乙酰化FOXO1的表达,并显著增强了Sal B诱导的SIRT3含量和对FOXO1去乙酰化的调节。SIRT3 siRNA转染后,Sal B诱导的乙酰化FOXO1下调被阻断,表明Sal B对细胞保护作用是通过SIRT3介导FOXO1去乙酰化所致。SIRT3/FOXO1通路是控制NAFLD氧化应激的关键治疗靶点,Sal B通过SIRT3介导FOXO1去乙酰化能减轻OA诱导的肝脂肪变性和氧化应激反应,对NAFLD细胞有保护作用。
基金
supported by grants from Wuhan Municipal Health and Family Planning Commission scientific research project(Grant No.WZ20Z04).
