Effect of TGF-βRⅡ siRNA on AngⅡ induced proliferation of glomerular mesangial cell / 实用医学杂志

ABSTRACT

Objective To investigate the effect of transforming growth factor β receptor-Ⅱ(TGF-βRⅡ) on AngⅡ inducing glomerular mesangial cell(GMC) proliferation and the expressions of TGF-β1 and psmad3 through transfecting TGF-βRⅡsmall interfering RNA(siRNA)into GMC. Methods Through transfecting fluo-rescence control siRNA into GMC ,we observed the transfection efficiency under fluorescence microscope after 6 hours. Transfecting TGF-βRⅡsiRNA and negative control siRNA into GMC respectively ,we detected the expres-sion of TGF-βRⅡ by western blot after 24 hours. The cells were divided into four groups:control group,AngⅡgroup,negative siRNA control group and TGF-βRⅡ siRNA group. Each group was stimulated by AngⅡ except the control group. After 24 hours,we detected the TGF-β1 and psmad3 protein levels by western blot and detected GMC proliferation by CCK8 kit. Results (1) Comparing with the scrambled control group,the expression of TGFβRⅡin the TGF-βRⅡsiRNA group was significantly decreased(P<0.05).(2)AngⅡcould accelerate the expression of TGF-β1. Transfecting TGF-βRⅡsiRNA into GMC decreased the expression of TGF-β1 protein(P<0.05).(3)AngⅡ could stimulate the phosphorylation of smad3. Transfecting TGF-βRⅡ siRNA into GMC de-creased the expression of psmad3 protein(P < 0.05).(4)Transfecting TGF-βRⅡ siRNA into GMC relieved the GMC proliferation AngⅡ-promoted(P < 0.05). Conclusions The AngⅡ stimulates the GMC proliferation,de-pending on the expression of TGF-βRⅡ. It is related to the expressions of TGF-β1 and psmad3.