高糖刺激大鼠肾间质成纤维细胞TGF-β、CTGF及α-SMA作用研究

Effects of TGF-β、CTGF and α-SMA on the NRK49f cultured 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(CTGF)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、24h及48h,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NRK49f12h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24h及48h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12h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NRK49f12h即可见TGF-β、CTGF mRNA表达均增加,至高糖刺激培养细胞24h及48h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NRK49f24h时可见α-SMA表达条带,至高糖DMEM培养48h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CTGF的转录和翻译,并可刺激其表达α-SMA。

Objective To detect the effects of TGF-β,CTGF andα-SMA on the NRK49 fcultured by high glucose.Methods The renal interstitial fibroblasts strain NRK49 fwas cultured with high glucose DMEM for 12 h,24 hand 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 andα-SMA expression.Results Compared with the control group,the TGF-βand CTGF expression were increased in the 24 hand 48 hhigh glucose group（P〈0.05）;the TGF-β,CTGF mRNA expression in the 24 hand 48 h high glucose group were higher than that in the control group and the 12 hhigh glucose group（P〈0.05）;theα-SMA expression were increased in the 24 hand 48 hhigh glucose group while not in the control and 12 hhigh glucose group.Conclusion The renal interstitial fibroblasts can increase the TGF-β,CTGF andα-SMA expression when stimulated by high glucose.