大黄酸对高糖诱导人肾小球系膜细胞Wnt、β-catenin蛋白及TGF-β1表达的影响

Effects of rhein on expressions of Wnt、β-catenin protein and TGF-β1 in high glucose-induced human mesangial cell

目的观察高糖对人肾小球系膜细胞Wnt、β-catenin蛋白表达、TGF-β1分泌的影响及大黄酸的干预作用。方法体外培养人肾小球系膜细胞.同步化后采用加有高浓度葡萄糖(30 mmol/L)联合不同浓度大黄酸的无血清培养基培养。应用MTT法测量高糖(HG组)及大黄酸高浓度组(HG+R1组)、大黄酸中浓度组(HG+R2组)、大黄酸低浓度组(HG+R3组)干预后肾小球系膜细胞增殖活力。各组细胞中Wnt、β-catenin蛋白采用放射免疫法检测。各组细胞上清液中转化生长因子β1(TGF-β1)采用ELISA法检测。结果①对系膜细胞增殖的影响:与正常对照组(NG)组24、48、72 h比较.HG组、HG+R1组、HG+R2组、HG+R3组细胞增殖均明显上升,差异均有统计学意义(P<0.05或P<0.01);与HG组各时点相比.HG+R1组、HG+R2组、HG+R3组在24 h呈下降趋势.48、72 h下降趋势显著(P<0.05或P<0.01),表现为时间、浓度依赖性;HG+R2组在24、48 h细胞增殖较HG+R3组各时点有下降趋势.72h时细胞增殖下降趋势显著(P<0.05).而HG+R1组细胞在48h、72h时细胞增殖均显著下降(P<0.05);与相比,HG+R1组细胞增殖抑制作用在72h时较HG+R2组显著下降(P<0.05)。②NG组细胞胞浆中有少量Wnt、β-catenin表达,HG组胞核表达Wnt胞浆及β-catenin量明显增高(P<0.05).大黄酸干预组Wnt、β-catenin蛋白表达受到抑制(P<0.05)。③细胞分泌TGF-β1受高糖刺激增加,TGF-β1的分泌受大黄酸抑制,均呈现出浓度依赖性(.P<0.05或P<0.01)。结论大黄酸在体外可能抑制Wnt、β-catenin蛋白表达.从而抑制分泌TGF-β1.

Objective To discuss theexpressions of Wnt、β-catenin protein and TGF-01 in high glucose-induced human mesangial cell and the intervention of rhein on it.Methods High concentration of glucose(30 mmol/L)combined with different concentrations of rhein were used to intervene cultured human mesangial cells.The activity of mesangial cell proliferation after all the interventions was examined by MTT measurement.Immunohistochemistry and ELISA were used to examine the expression of Wnt、β-catenin protein and TGF-β1.in normal mesangial cells and cells intervened by high glucose and Rhein.Results ①Inhibition effect to human mesangial cells:compared with NG group for 24 h.48 h and 72 h,human mesangial cells proliferation in HG group,HG+R1 group,HG+R2 group,and HG+R3 group were increased significantly(P<0.05 or P<0.01);Compared with HG group for 24 h.48 h and 72 h.the HG+R1 group,the HG+R2 group and the HG+R3 group at 24 h showed a downward trend;and the trend de creased significantly at 48 h and 72 h(P<0.05 or P<0.01),and the performance expressed a time,concentration-dependent.Compared with HG+ R3 group for 24 h.48 h and 72 h.the HG+R2 group showed a downward trend at 24 h.48 h.but the trend decreased significantly at 72 h(P<0.05).The HG+R1 group showed a downward trend at48 h.72 h(P<0.05).Compared with HG+R2 group for 24 h.48 h and 72 h.the HG+R1 group at 72 h showed a downward trend(P<0.05).②The mesangial cells expressed Wnt and β-catenin at a certain amount level in the normal state.The expression of Wnt、β-eatenin protein increased when they were stimulated by high glucose,(P<0.05);while the expression of them was significantly reduced in high glucose-induced mesangial cells in HG+R1 group.HG+R2 group and HG+R3 group(P<0.05).③Cells secrete TGF-β1 was promoted by the high glucose.The secretion of TGF-β1 was decreased in HG+R group(P<0.05 or P<0.01).Conclusion The proliferation of human mesangial cells would be inhibited by rhein in vitro,which were promoted by hyperglycemia,the down- regulating expression of Wnt,β-catenin protein and the secretion of TGF-β1 may be related to the mechanism of which.