Ring Spore, A (csa) Impact On Cultivating Human Mesangial Cells Proliferation In Vitro Mechanism Research

Objective1To investigate the effect of CsA on proliferation,cell cycle of human mesangial cells and expression of cyclinDl,cyclinE, p27kipl protein in human mesangial cells.2To study the mechanism of CsA on proliferation of human mesangial cells.MethodsRecover the frozen mesangial cell in37℃water bath snd move it into50ml culture bottles,then it shuld be placed in37℃,5%CO2incubator.When adherent cells up to80%,passage the cells and continue to culture,spread3-4generations,the cell viability was enough strong for the experiments.Divided into experimental and control groups,experimental group with different concentrations of CsA(0.1,1,5μmol/L),the control group without CsA-treated.Measngial cells were inoculated on culture plates.The first step is:Human mesangial cells were added different concenteations of CsA,measured proliferation inhibition of HMC with MTT at24h,48h and72h at the490nm wavelength.The second step is:Human mesangial cells were added different concenteations of CsA,measured cell cycle of mesangial cells with FCM after48h.The third step is:Add different concenteations of CsA in human mesangial cells, after48h.the expression of cyclinDl1、cyclinE and p27kipl proteins in cell nucleus was assessed by Westorn blot assay.Results1The proliferation of human mesangial cells Was inhibited in a time-and dose-dependent manner by CsA (P<0.05)2The cell cycle was arrested in G0/G1phase. The percentage of G0/G1phase cells gradually increased with the rising of CsA concentrations (P<0.05)3In cell nucleus the expression of cyclinDl.cyclinE proteins decreased and p27kipl protein increased significantly with the rising concentrations of CsA (P<0.05).Conclusion1CsA can orderly block the cell cycle, significantly inhibit the proliferation of cultured human mesangial cells in vitro.2The expression of cyclinDl,cyclinE proteins decreased and p27kipl protein increased significantly in cell with the rising concentrations of CsA (P<0.05). 3The mesangial cell proliferation could be inhibited by CsA, the mechanism may be related to the decrease of cyclinD1,cyclinE proteins and the increase of p27kip1protein..