大黄酸对高糖所致肾小球系膜细胞Wnt／β-catenin信号通路的影响探讨

Effect of rhein on high glucose-induced mesangial cell Wnt-β-catenin Signaling Pathway

目的观察高糖对人肾小球系膜细胞（HMC）Wnt／β-catenin信号通路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（30mmol／1）联合不同浓度大黄酸的无血清培养基培养。应用四甲基偶氮唑蓝（MTT）测量高糖（HG组）及大黄酸高浓度组（高糖＋大黄酸10^-4mol／L，HG＋R1组）、大黄酸中浓度组（高糖＋大黄酸10^-5mol／L，HG＋R2组）、大黄酸低浓度组（高糖＋大黄酸10^-6mol／L，HG＋R3组）干预后肾小球系膜细胞增殖活力。应用RT-PCR法检测正常培养及高糖作用后人肾小球系膜细胞Wnt／β-catenin基因的表达及大黄酸对其表达的影响。结果①对系膜细胞的抑制作用：与正常对照组（NG）组24h、48h、72h（OD值分别为0．169±0．051、0．228±0．074、0．285±0．075）比较，HG组（OD值分别为0．307±0．074、0．507±0．038、0．711±0．075）、HG＋R1组（OD值分别为0．241±0．027、0．334±0．015、0．499±0．063）、HG＋R2组（OD值分别为0．244±0．081、0．386±0．033、0．531±0．011）、HG＋R3组（OD值分别为0．277±0．036、0．407±0．057、0．594±0．042）细胞增殖均明显上升，差异均有统计学意义（P〈0．05、P〈0．01）；与HG组各时点相比，HG＋R1组、HG＋R2组、HG＋R3组在24h呈下降趋势，48h、72h下降趋势显著，表现为时间、浓度依赖性（P〈0．05、P〈0．01）。②在基础状态下，系膜细胞表达一定量的Wnt／β-catenin经高糖刺激后，Wnt／β-catenin基因表达上调（P〈0．05），HG＋R1组、HG＋R2组、HG＋R3组均可明显下调高糖刺激引起的系膜细胞Wnt／β-catenin基因表达（P〈O．05）。结论大黄酸可能通过下调Wnt／β-catenin基因的表达抑制高糖引起的HMC增殖。

Objective To discuss the effects of high glucose on human mesangial cells （HMC） Wnt-β-catenin signaling pathway, and the intervention of rhein on it. Methods High concentration of glucose （30 mmol/L） combined with different concentrations ofrhein were used to intervene cultured human mesangial cells. The activity of mesangial cell proliferation after all the interventions was examined by MTT measurement. Total Wnt~ β-catenin RNA was detected By RT-PCR in normal mesangial cells and cells intervened by high glucose and Rhein. Results （1）Inhibition effect to human mesangial cells： compared with NG group for 24 h, 48 h and 72 h （OD values were 0.169±＋0.051, 0.228±0.074, 0.285±0.075）, human mesangial cells proliferation in HG group （OD values were 0.307± 0.074, 0.507 q-0.038, 0.711± 0.075）, HGq-R1 group （OD value were 0.241 ± 0.027, 0.334 4± 0.015, 0.499 ± 0.063）, HGq-R2 group （OD value were 0.244± 0.081, 0.386 ± 0.033, 0.531±0.011）, and HG-R3 group（OD value were 0.277±0.036, 0.407±0.057, 0.594±0.042） were increased significantly（P〈0.05, P〈0.01） ; Compared with HG group for 24 h, 48 h and 72 h, the HGq-R1 group, the HGq-R2 group and the HGq- R3 group showed a downward trend at 24 h, but not significantly; but the trend decreased significantly at 48 h and 72 h, and the performance expressed a time, concentration-dependent（P〈0.05, P〈0.01）. （g） In the normal state, the mesangial ceils expressed a certain amount of Wnt and β-catenin, when they were stimulated by high glucose, the expression of Wnt, β-catenin in mRNA increased （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）. Conclusion Rhein may inhibit the proliferation of high glucose-induced mesangial cells through the Wnt and β-catenin gene expression.