基于SIRT1/SIRT3/FOXO1信号通路探讨染料木素减轻酒精性脂肪肝的作用机制OA

Objective To investigate whether genistein alleviates alcoholic fatty liver disease(AFLD)by regulating sirtuin1(SIRT1)/sirtuin3(SIRT3)/forkhead box protein O1(FOXO1)signaling pathway.Methods A HepG2 AFLD model was induced using 100 μmol/L oleic acid combined with 150 mmol/L 95%ethanol.The experiment was divided into 16 groups,including control,model,genistein,and resveratrol groups,SIRT1 gene silencing control+model,genistein,and resveratrol groups(C-shSIRT1,M-shSIRT1,G-shSIRT1,and R-shSIRT1,respectively),SIRT3 gene silencing control+model,genistein,and resveratrol groups(C-shSIRT3,M-shSIRT3,G-shSIRT3,and R-shSIRT3,respectively),and SIRT1/SIRT3 dual gene silencing control+model,genistein,and resveratrol groups(C-shSIRT1/3,M-shSIRT1/3,G-shSIRT1/3,and R-shSIRT1/3,respectively).Cellular steatosis was analyzed through Oil Red O staining and measurements of triglyceride,total cholesterol,and free fatty acid levels.Intracellular levels of tumor necrosis factor-α and interleukins 6 and 1β were determined by enzyme-linked immunosorbent assay.Western blot was used to detect protein expression levels of SIRT1,SIRT3,and FOXO1,and their interactions were analyzed by co-immunoprecipitation.Results Genistein significantly reduced lipid deposition and inflammatory responses in HepG2 AFLD model cells(P<0.05).Compared with the control group,protein expression of SIRT1,SIRT3,and FOXO1 was significantly decreased in the model group(P<0.01).Compared with the model group,the genistein group showed significantly increased protein expression of SIRT1,SIRT3,and FOXO1(P<0.05).Co-immunoprecipitation result showed that SIRT1,SIRT3 and FOXO1 interacted.Genistein significantly reduced cellular steatosis after either SIRT1 or SIRT3 gene silencing,but its anti-AFLD effect was attenuated when both genes were simultaneously silenced.Compared with the C-shSIRT1 group,FOXO1 expression was significantly increased in the M-shSIRT1 group(P<0.01).Compared with the M-shSIRT1 group,FOXO1 expression was significantly decreased in the G-shSIRT1 group(P<0.01).Compared with the C-shSIRT3 group,FOXO1 expression was significantly decreased in the M-shSIRT3 group(P<0.01).Compared with the M-shSIRT3 group,FOXO1 expression was significantly increased in the G-shSIRT3 group(P<0.01).Compared with the C-shSIRT1/3 group,FOXO1 expression was significantly decreased in the M-shSIRT1/3 group(P<0.01);FOXO1 expression levels were similar between the M-shSIRT1/3 and G-shSIRT1/3 groups(P>0.05).Conclusions Genistein ameliorates lipid metabolism and suppresses inflammatory response in HepG2 AFLD cells by regulating SIRT1/SIRT3/FOXO1 signaling pathway,with the SIRT3/FOXO1 axis playing a particularly crucial role.