Regulation Of Adenovirus Type 36 For PI3K/AKT Signaling Pathway

Objective:To investigate the effect of Ad36 to PI3 K / AKT signaling pathway downstream of PPAR? and lipid metabolic regulation of related genes during human adipose-derived mesenchymal stem cells differentiate into adipocyte induced by Ad36.Methods:(1)Induce hAMSC with adenovirus,RT-qPCR detect the expression of adenovirus E1 A gene,oil red O staining adipocytes,Western Blotting detect PPAR? of adipocyte,identificate Ad36-induced adipocyte differentiation model.(2)Determinate culture medium glucose concentration and intracellular triglyceride content.(3)RT-qPCR,Western Blotting detects FoxO1?LPIN1?APM1?ACC?GLUT4 genes expression levels of adipocytes.(4)Chromatin Immunoprecipitation experiments detect FoxO1 and PPAR?regulation to its target genes.(5)After suppressing the expression of P-FoxO1 with Wortmannin,Western Blotting detected PPAR? protein expression levels,RT-qPCR detected LPIN1,APM1,ACC,GLUT4 genes expression levels in Ad36-induced human adipocytes.Results:(1)Ad36 induced hAMSC differentiate to adipocytes,adipocyte differentiation marker of PPAR? mRNA and protein expression levels significantly higher than the control group(P<0.05),the process of differentiation medium glucose levels decreased significantly than the control group(P<0.05).(2)Ad36 induced hAMSC differentiate to hAMSC,LPIN1,APM1,ACC,GLUT4 genes expression level was significantly higher compared with the control group(P<0.05),FoxO1 protein expression levels were significantly decrese after Ad36 induced hAMSC(P<0.05),P-FoxO1 protein expression levels increased significantly after Ad36 induced hAMSC(P<0.05).(3)The PPAR? promoter region presence of FoxO1 binding site,FoxO1 binding to the promoter region of PPAR?,inhibate PPAR? transcriptional activity.After using Wortmannin to inhibite the expression of P-FoxO1,PPAR? protein expression levels significantly reduced.(4)LPIN1,APM1,ACC,GLUT4 genes promoter region presence of PPAR? binding sites,PPAR? activate gene expression levels by combine with its promoter region.In using wortmannin,the fat cells gene expression of LPIN1,APM1,ACC,GLUT4 levels were significantly decresed(P<0.05).Conclusion:Ad36 raised lipid metabolism genes of LPIN1?APM1?ACC?GLUT4 expression levels by increasing downstream of PI3K/AKT signal pathway of FoxO1 and PPAR? and promotes adipocyte differentiation..