| Objective To investigate the effects of ERK1/2 and leflunomide intervention in podocyte induced by high glucose and relevant mechanisms.Methods human podocytes cultured in vitro were divided into normal glucose group, high permeability group, high glucose group which was detected at 0.5h,1h,6h,12h,24h after induced. Changes of ERK1/2 signal pathway in each group were detected by using Western blotting. Detection of podocyte apoptosis: minutes, reached the peak at hour 6; maintained the activation to hour 24, and than declined to the basal level. However, activation of ERK1 /2 was not detected in normal glucose and high podocytes were divided into high glucose control group,inhibitor group, inhibitor+ A771726 group, and A771726 group. Changes of ERK1/2 signal pathway and podocyte apoptosis in each group were detected by using Western blotting and flow cytometry respectively after cultured for 24 hours.Results(1) High glucose medium induced phosphorylation of ERK1/2 as early as 30 permeability groups.(2) Compared with high glucose group, phosphorylation of ERK1/2 was blocked by PD98059, a specific ERK1/2 activation inhibitior,which attenuated the high glucose-induced expression of P-ERK1/2 obviously(P=0.003)and result to more podocyte apoptosis by influencing the activation of many downstream nuclear transcription factors and expression of target gene(P=0.001).(3) A771726 reduced podocyte apoptosis(P=0.021) by increasing expression of P-ERK1/2 (P=0.002).However, PD98059 influencd the protection of A771726 to podocytes and result to more apoptosis compared with high glucose group(P=0.006).Conclusion ERK1/2 signal pathway takes part in the protection of Leflunomide to podocytes. |
