丹酚酸B对非酒精性脂肪肝细胞模型SIRT1/SIRT3信号通路的研究  

Study on the influence of salvianolic acid B on vSIRT1/SIRT3 signaling pathway in non-alcoholic fatty liver cell model

作　　者：任振晓 张佳佳[2] Ren Zhenxiao;Zhang Jiajia(Qilu Medical University,Zibo,Shandong,255300,China;Department of Pharmacy,Women’s Hospital of Nanjing Medical University(Nanjing Maternity and Child Health Care Hospital,Nanjing,Jiangsu,210004,China)

机构地区：[1]齐鲁医药学院,山东淄博255300 [2]南京医科大学附属妇产医院(南京市妇幼保健院)药学部,210004

出　　处：《齐齐哈尔医学院学报》2022年第24期2313-2318,共6页Journal of Qiqihar Medical University

摘　　要：目的检测丹酚酸B(Salvianolic acid,Sal B)是否可以通过影响SIRT1/SIRT3信号通路对棕榈酸(Palmitic acid,PA)诱导AML-12细胞建立的非酒精性脂肪肝(non-alcoholic fatty liver disease,NAFLD)细胞模型起到保护作用。方法将AML-12细胞随机分为五组:正常对照组、Sal B(16μM)给药组、PA组、PA+Sal B(2μM、16μM)预处理组。采用试剂盒检测法测量谷丙转氨酶(Alanine transaminase,ALT)、谷草转氨酶(Alanine transaminase,AST)、甘油三酯(triacylglycerol,TG)、总胆固醇(Total cholesterol,TC)、丙二醛(malonaldehyde,MDA)、超氧化物歧化酶(superoxide,SOD)含量;Western Blot法检测细胞内SIRT3、SIRT1蛋白的表达qRT-PCR法检测细胞内SIRT3mRNA、SIRT1mRNA的表达。另外,对SIRT1进行RNA干扰实验,分为正常对照组、PA组、PA+Sal B预处理组、PA+SIRT1 siRNA预处理组、PA+Sal B+SIRT1 siRNA预处理组五组。采用Western Blot法及qRT-PCR法检测细胞内SIRT3、SIRT1蛋白表达及mRNA水平。其中PA+Sal B+SIRT1 siRNA预处理组方法为先进行SIRT1 siRNA转染48 h,然后给予Sal B(16μM)预处理6h,最后用PA(200μM)诱导24 h。结果PA组较对照组ALT、AST、TC、TG、MDA水平明显升高(P<0.01),SOD活力显著下降(P<0.01);Sal B组较PA组ALT、AST、TC、TG、MDA水平明显降低(P<0.01),SOD活力明显升高(P<0.01)。另外,PA组较对照组SIRT1和SIRT3蛋白表达水平及mRNA水平显著下降(P<0.01);Sal B组较PA组SIRT1和SIRT3蛋白表达水平及mRNA水平显著升高(P<0.01)。PA+SIRT1 siRNA组较PA组SIRT3蛋白表达水平明显降低(P<0.01),Sal B+PA+SIRT1 siRNA组较PA组SIRT3蛋白表达水平未升高。结论Sal B通过上调SIRT1/SIRT3信号通路改善PA诱导的AML-12细胞非酒精性肝损伤。Objective To test whether salvianolic acid B(SalB)can play a protective role on the palmitic acid(PA)induced AML-12 cells model of non-alcoholic fatty liver(NAFLD)by affecting the SIRT1/SIRT3 signaling pathway.Methods AML-12 cells were randomly divided into five groups:normal control group,Sal B(16μM)administration group,PA group,and PA+Sal B(2μM,16μM)preprocessing group.The contents of alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG),total cholesterol(TC),malondialdehyde(MDA)and superoxide dismutase(SOD)were measured by kit detection method.Western Blot method was used to detect the expression of intracellular SIRT3 and SIRT1 proteins and qRT-PCR method was used to detect intracellular expression of SIRT3mRNA and SIRT1mRNA.In addition,RNA interference experiments were performed on SIRT1,which were divided into five groups:normal control group,PA group,PA+Sal B pretreatment group,PA+SIRT1 siRNA pretreatment group,and PA+Sal B+SIRT1 siRNA pretreatment group.Western Blot method was used to detect the protein expression and mRNA level of SIRT3 and SIRT1 in cells were detected by qRT-PCR.The PA+Sal B+SIRT1 siRNA pretreatment group was firstly transfected with SIRT1 siRNA for 48 h,then pretreated with Sal B(16μM)for 6 h,and finally induced with PA(200μM)for 24 h.Results Compared with control group,the levels of ALT,AST,TC,TG and MDA in PA group were significantly increased(P<0.01),however,SOD activity was significantly decreased(P<0.01).Compared with PA group,the levels of ALT,AST,TC,TG and MDA in Sal B were significantly decreased(P<0.01),the activity of SOD was significantly increased(P<0.01).Compared with the control group,the protein expression levels and mRNA levels of SIRT1 and SIRT3 in PA group were significantly decreased(P<0.01).The protein expression levels and mRNA levels of SIRT1 and SIRT3 in Sal B group were significantly higher than those in PA group(P<0.01).The SIRT3 protein expression level in the PA+SIRT1 siRNA group was significantly lower than that in the PA group(

关 键 词：丹酚酸B SIRT3 SIRT1 非酒精性脂肪肝 

分 类 号：R575.5[医药卫生—消化系统]