摘要目的 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(30 mmol/l)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(高糖十大黄酸10-4 mol/L,HG+R1组)、大黄酸中浓度组(高糖+大黄酸10-5 mol/L,HG+R2组)、大黄酸低浓度组(高糖十大黄酸10-6 mol/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、48 h、72 h (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、0334±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24 h呈下降趋势,48 h、72 h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＜0.05).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.

abstractsObjective 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 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.Total Wnt、β-catenin RNA was detected By RT-PCR 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 (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 ±0.038,0.711±0.075),HG+R1 group(OD value were 0.241± 0.027,0.334±0.015,0.499±0.063),HG+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 iucreased significantly (P＜0.05、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 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).② In the normal state,the mesangial cells expressed s 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.