Salvianolic acid B protects against hypoxia/reoxygenation-induced renal tubular epithelial cells damage via regulating AMPK mediated ferroptosis
Objective:To investigate the protective effects and mechanisms of salvianolic acid B(Sal B)on hypoxia/reoxygen-ation(H/R)-induced renal tubular epithelial cell damage.Methods:In vitro experimental cells were divided into the normal con-trol(Con)group,the Sal B+Con group,the H/R group,the Sal B+H/R group,as well as the Con group,the H/R group,Sal B+H/R group,the H/R+CC group,and the H/R+Sal B+CC group.HK-2 cell viability was detected by the CCK-8 method.Cell lysates were collected to measure the levels of GSH and malondialdehyde MDA using an enzyme-linked immunosorbent as-say.The expressions of p-AMPK,GPX4,ACSL4 and FSP1 were detected by Western blot.The expression of ferroptosis mark-ers GPX4 and ACSL4 proteins was detected by cell immunofluorescence,while mitochondrial membrane potential and cell apopto-sis were measured by flow cytometry.Results:The results of the in vitro experiments showed that Sal B had no significant toxicity on normal HK-2 cell viability within the concentration range of 20-160 μmol/L(P>0.05).Compared with the model group,Sal B at concentrations of 20,40,and 80 μmol/L dose-dependently increased the viability of HK-2 cells(P<0.01).Compared with the model group,MDA content and GSH content of the H/R+Sal B group were decreased,p-AMPK,GPX4 and FSP1 protein expression levels were increased,ACSL4 protein expression was decreased,mitochondrial membrane potential was significantly increased,and apoptosis rate was significantly decreased(P<0.05).Compared with the H/R+Sal B group,the FSP1 and GPX4 protein expression level in the H/R+AMPK group was decreased,the ACSL4 protein expression level was increased(P<0.05),and the mitochondrial membrane potential was significantly decreased(P<0.05),and the addition of Sal B could reverse this situation(P<0.05).Conclusion:Salvianolic acid B can alleviate I/R-induced AKI,and its mechanism may involve the activa-tion of the AMPK pathway to mitigate ferroptosis in renal tubular epithelial cells.
万方数据
维普
