Effect of knockdown of PRDX6 on adaptive expression of bile acid transporter in HepG2 cells induced by rifampicin / 安徽医科大学学报

ABSTRACT

Objective @#To investigate the role of knockdown of peroxiredoxin-6 ( PRDX6) in injury and adaptive expression of bile acid transporter in human hepatoellular carcinomas ( HepG2 ) cells induced by rifampicin (RFP) . @*Methods @#Cells in logarithmic growth phase were uniformly inoculated in six-well plates , and HepG2 cells were transiently transfected with specific PRDX6-siRNA and control-siRNA to construct the knockdown group and control group . After 24 h of induction with 100 μmol/L RFP , Western blot and qRT-PCR were performed to detect the protein and gene expression levels of PRDX6 , multidrug resistance protein 1 (MDR1) , multidrug resist- ance-associated proteins 2 , 3 and 4 (MRP2 , MRP3 and MRP4) , and Na + /taurine taurocholate cotransporter pro- tein (NTCP) . Annexin V-FITC/PI double staining assay was used to detect the apoptosis rate of cells in each group ; CCK-8 assay was used to detect the changes of cell proliferation in each group; The relative contents of ala- nine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , total bilirubin ( TBIL) , indirect bilirubin (IBIL) and total bile acid (TBA) in the supernatant of cell culture medium of each group were detected by kits .@*Results @#RFP increased the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 , NTCP and PRDX6 in HepG2 cells (P < 0. 05) , while the protein and gene expression levels of MRP2 , MRP3 , MRP4 , MDR1 and NTCP decreased to different degrees after PRDX6 knockdown (P < 0. 05) . In addition , PRDX6 knockdown re- sulted in increased apoptosis rate of HepG2 cells (P < 0. 05) , decreased cell proliferation ability (P < 0. 05) , and increased levels of cell injury markers (ALT , AST , TBIL , DBIL , TBA) in cell culture supernatants (P < 0. 05) . @*Conclusion @#RFP increased the protein and gene expression of bile acid transporter and PRDX6 to increase in HepG2 cells . However , following knockdown of PRDX6 and treatment with RFP , the protein and gene expression levels of the bile acid transporter decreased and cell injury was aggravated , suggesting that PRDX6 played a protec- tive role in RFP-induced adaptive response in HepG2 cells .