The Role Of TRPC6 In High Glucose Induced Podocytes Injury

Objective:To explore the effect of high glucose on the expression of TRPC6 in podocytes cultutred in vitro and investigate the role of TRPC6 in high glucose induced podocytes injury.Methods:Immortalized mouse podocyte cell line was cultured and divided into three groups:control group, treated with normal concentration of D-glucose(5.6mmol/L),HG group, treated with high concentration of D-glucose(30mmol/L),and mannitol group,treated with mannitol (25mmol/L) plus D-glucose(5.6mmol/L).Then shRNA targeting TRPC6 gene (PGCsi3.0-TRPC6B shRN)was transfected to podocytes by liposome. The level of TRPC6,desmin,nephrin mRNA were detected by RT-PCR;the level of TRPC6,desmin,nephrin proteins were detected by Western blot analysis;the distribution and expression of F-actin was detected by laser scanning confocal microscopy;the apoptosis rate of podocytes was detected by hoechst33258.Results:An up-regulation of mRNA and protein expression of TRPC6,desmin and down regulation of mRNA and protein expression of nephrin were detected in HG group compared with control group. High glucose ambient also decreased the ratio of podocytes with distinct F-actin fibers and increased the apoptosis rate. Transfection with PGCsi-TRPC6B shRNA down-regulated expression of TRPC6,desmin and up-regulated expression of nephrin, increased the ratio of podocytes with distinct F-actin fibers and decreased the apoptosis rate.Conclusion:High glucose may increase the expression of TRPC6 in podocytes.The alteration of expression of TRPC6 may participate in the regulation of slit diaphragm and cytoskeleton in high glucose ambient.