Protective Effect Of Pancreatic Kininogenase On Human Kidney Proximal Tubular Epithelial Cell Injury Induced By Hydrogen Peroxide

Objective:To investigate the protective effect of pancreatic kininogenase(PK)on human kidney proximal tubular epithelial(HK-2)cells injury induced by hydrogen peroxide(H2O2).Methods:HK-2 cells lines were cultured in vitro and randomly divided into 4 groups:control(CON)group,H2O2(500uM)group,PK(6pg/mL)group and H2O2+ PK group.The treatment time of each group was 24h.The cell viability of each group was determined by the CCK-8;The production of intracellular reactive oxygen species(ROS)was detected by DCFH-DA fluorescence coupled with flow cytometry;The apoptosis rate was detected by flow cytometry;Western blotting was used to detect the expression of superoxide dismutase 2(SOD2),NADPH oxidase 4(NOX4),transforming growth factor-β1(TGF-β1),monocyte chemoattractant protein-1(MCP-1),autophagy related factors light chain protein 3B(LC3B)and Beclin-l,b-cell lymphoma-2(Bcl-2)and bcl2-associated X(Bax).Results:Compared with the CON group,the viability of HK-2 cells decreased,the generation of ROS increased,the expression of SOD2 protein decreased,the expression of NOX4,TGF-β1,MCP-1,LC3B-Ⅱ/LC3B-Ⅰ,Beclin-1 protein and the generation of apoptosis rate increased,the expression of Bcl-2/Bax protein decreased in the H2O2 group(P<0.001);Compared with the H2O2 group,the viability of HK-2 cells increased,the generation of ROS decreased,the expression of SOD2 protein increased,the expression of NOX4,TGF-β1,MCP-1,LC3B-II/LC3B-I,Beclin-1 protein and the generation of apoptosis rate decreased,the expression of Bcl-2/Bax protein increased significantly in the H2O2+PK group(P<0.01).All had statistical significance.Conclusion:PK has a protective effect on HK-2 cells injury induced by H2O2,the inhibition of TGF-β1 expression,anti-oxidative stress,regulation of autophagy and anti-apoptosis.