Tacrolimus inhibits Puromycin induced injury of podocytes by stabilizing the expression and distribution of podocin / 中华实用儿科临床杂志

ABSTRACT

Objective To investigate the effect of Puromycin(PAN)and Tacrolimus(FK506)on the expre-ssion of podocin in podocytes,and discuss the mechanism of FK506 in improving proteinuria caused by the damage of glomerular podocytes.Methods Mouse podocyte were cultured and divided into 3 groups,which were control group, PAN group and FK506 group,respectively.The changes of each group after 8 hours,24 hours and 48 hours of treatment were detected by using phase-contrast microscope,and the expression and distribution of protein and mRNA were tested by adopting Western blot,quantitative real-time PCR and immunofluorescence technique.Results Compared with the control group(8.54 ± 0.25,8.27 ± 0.07,7.45 ± 0.13)at each time point(8 hours,24 hours and 48 hours),the soma size of the PAN group(6.41 ± 0.22,4.96 ± 0.09,3.76 ± 0.16)was reduced.But in the FK506 group(7.67 ± 0.06, 6.62 ± 0.04,5.57 ± 0.27),it was increased at each time point(8 hours,24 hours and 48 hours)compared with the PAN group.The podocytic process and the intercellular connection disappeared,and the distribution was significantly scattered.The mRNA(0.87 ± 0.15,0.78 ± 0.15,0.58 ± 0.12)and protein(0.82 ± 0.02,0.62 ± 0.03,0.50 ± 0.02) expressions of podocin increased in FK506 group and mRNA(0.63 ± 0.12,0.56 ± 0.01,0.48 ± 0.02),protein (0.71 ± 0.03,0.46 ± 0.01,0.34 ± 0.02)were observably reduced in PAN group at each time point(8 hours,24 hours and 48 hours)and showed abnormal distribution in PAN group,compared with the control group[podocin mRNA (1.22 ± 0.15,1.18 ± 0.06,0.87 ± 0.30),protein(0.86 ± 0.03,0.87 ± 0.03,0.61 ± 0.07)].Conclusions PAN attenuates the mRNA and protein expression of podocin by damaging podocytes,inversely,while FK506 protects poto-cyte injury by stabilizing the mRNA and protein expression of podocin,which maybe involve inhibition of proteinuria.It can be used as a target for the study and treatment of kidney diseases.