高糖刺激大鼠肾间质成纤维细胞TGF-β、CTGF及α-SMA作用研究

摘要

Objective To detect the effects of TGF-β,CTGF and α-SMA on the NRK49f cultured by high glucose.Methods The renal interstitial fibroblasts strain NRK49f was cultured with high glucose DMEM for 12 h,24 h and 48 h,the normal DMEM media was used as control.Western Blot and real-time RT-PCR were applied to analyze the TGF-β,CTGF and α-SMA expression.Results Compared with the control group,the TGF-β and CTGF expression were increased in the 24 h and 48 h high glucose group (P＜0.05);the TGF-β,CTGF mRNA expression in the 24 h and 48 h high glucose group were higher than that in the control group and the 12 h high glucose group (P＜0.05);the α-SMA expression were increased in the 24 h and 48 h high glucose group while not in the control and 12 h high glucose group.Conclusion The renal interstitial fibroblasts can increase the TGF-β,CTGF and α-SMA expression when stimulated by high glucose.%目的 探讨高糖对大鼠肾间质成纤维细胞株NRK49f转化生长因子-β(TGF-β)、结缔组织生长因子(CT-GF)及α-平滑肌肌动蛋白(α-SMA)表达的影响.方法 将处于对数生长期的大鼠肾间质成纤维细胞株NRK49f用无血清高糖DMEM培养12h、24 h及48 h,同时设空白对照组,用Western Blot及实时定量RT-PCR法检测并分析各组细胞TGF-β、CTGF及α-SMA表达情况.结果 高糖DMEM作用于NRK49f 12 h即可见TGF-β、CTGF蛋白表达均有增加趋势,与正常对照组比较,高糖刺激24 h及48 h组TGF-β相对表达量均明显增加,差异有统计学意义(P＜0.05),且与高糖12 h组比较,二者相对表达量变化明显增加(P＜0.05);高糖DMEM作用于NRK49f 12 h即可见TGF-β、CTGF mRNA表达均增加,至高糖刺激培养细胞24 h及48 h时,二者相对表达量均明显高于对照组及高糖培养12h组,差异均有统计学意义(P＜0.05);高糖DMEM作用于NRK49f 24 h时可见α SMA表达条带,至高糖DMEM培养48 h时,α-SMA相对表达量最高.结论 高糖可刺激体外培养的肾间质成纤维细胞表达TGF-β和CT-GF的转录和翻译,并可刺激其表达α-SMA.