Effect of knockdown of PRDX6 on adaptive expression of bile acid transporter in HepG2 cells induced by rifampicin
10.19405/j.cnki.issn1000-1492.2024.04.016
- VernacularTitle:敲低PRDX6对利福平诱导HepG2细胞胆汁酸转运体适应性表达的影响
- Author:
Liufang QU
1
;
Mengxue HUANG
;
Shiguo CAO
;
Gang CHEN
;
Jianming XU
;
Weiping ZHANG
Author Information
1. 安徽医科大学第一附属医院消化内科,合肥 230022
- Keywords:
peroxiredoxin-6;
rifampicin;
bile acid transporter;
adaptive response;
cell injury;
apoptosis
- From:
Acta Universitatis Medicinalis Anhui
2024;59(4):660-665
- CountryChina
- Language:Chinese
-
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.
