Effect of decoy receptor 3 gene on hepatocyte apoptosis

Objective To investigate the effect of decoy receptor 3( DcR3) gene on hepatocyte apoptosis,as well as the possible mechanism of action of DcR3 in this process. Methods The human liver cell lines were cultured in vitro,and p EF1α- DcR3 transfection group,p EF1α- IRES transfection group,and negative control group were established. The p EF1α- IRES- DsRed- Express2- DcR3 eukaryotic expression vector was constructed and transfected into human liver cell lines for 36 hours. qRT- PCR was used to measure the mRNA expression of DcR3,Fas ligand( Fas L),α- smooth muscle actin( α- SMA),and transforming growth factor- β1( TGF- β1),Western blot was used to measure the change in the protein expression of DcR3,and flow cytometry was used to measure apoptosis. An analysis of variance was used for comparison of continuous data between groups,and the least significant difference t- test was used for comparison between any two groups. Results After human liver cell lines were transfected with p EF1α- DcR3 for 36 hours,the p EF1α- DcR3 transfection group showed significant increases in the mRNA and protein expression of DcR3 compared with the p EF1α- IRES transfection group and negative control group( F = 33 169. 5and 141. 54,all P < 0. 01). Compared with the other two groups,the p EF1α- DcR3 transfection group showed significant reductions in the mRNA expression of Fas L,α- SMA,and TGF- β1( F = 269 451. 8,20 790. 4,and 8067. 8,all P < 0. 01),which suggested that DcR3 inhibited the expression of Fas L,α- SMA,and TGF- β1. Compared with the p EF1α- IRES transfection group and negative control group,the p EF1α- DcR3 transfection group showed a significant reduction in apoptosis rate( F = 558. 63,all P < 0. 01). Conclusion DcR3 can inhibit hepatocyte apoptosis and downregulate the mRNA expression of Fas L,α- SMA,and TGF- β1.