虫草素通过下调TGF-β抑制高糖诱导的大鼠肾小管上皮细胞转分化

Cordycepin Inhibit Epithelial- mesenchymal- transition of NRK52E Cells Induced by High Glucose by Downregulate TGF-β Expression

目的:探讨虫草素对高糖诱导的大鼠肾小管上皮细胞转分化的影响。方法:体外培养大鼠近端肾小管上皮细胞株(NRK52E细胞株),分为正常对照组(葡萄糖5.5 mmol/L,NG组)、高糖组(葡萄糖30 mmol/L,HG组)、高糖+虫草素组(葡萄糖30 mmol/L+虫草素10μg/ml,HG+C组)。分别于刺激12 h,24 h,48 h后收集细胞。应用定量RT-PCR测定NRK52E TGF-β,E-cadherin,α-SMA mRNA的表达;Western印迹方法检测TGF-β、E-cadherin、α-SMA蛋白的表达。结果:高糖刺激后NRK52E细胞的TGF-β和α-SMA mRNA及蛋白表达明显高于正常糖组(P<0.01),而虫草素组TGF-β和α-SMA mRNA及蛋白表达显著低于高糖组(P<0.05);高糖诱导的NRK52E细胞E-cadherin mRNA及蛋白水平明显降低(P<0.01);而虫草素组NRK52E细胞E-cadherin mRNA及蛋白水平显著高于高糖组(P<0.05)。结论:虫草素可以明显抑制高糖诱导的大鼠肾小管上皮细胞转分化,其机制可能是通过下调TGF-β实现。

Objective：To explore the role of cordycepin on epithelial-mesenchymal-transition of NRK52E cells induce by high glucosed. Methods：NRK52E cells were cultured in the mediun with normal glucose concentration （glucose 5. 5 mmol/L, group NG） , high glucose concentration （ glucose 30 mmol/L, group HG ） and high glucose. Concentration with cordycepin （ glucose 30 mmol/L, and Cordycepin 10μg/ml group, HG and C）. Cells were collected in 12H, 24H and 48H after the stimulation of normal glucose, high glucose, and high glucose with cordycepin respectively. The expression of TGF-β, E-cadherin andα-SMA mRNA was measured by quantitative RT-PCR; The expression of TGF-β1 , E-cadherin and α-SMA protein was detected by western blot. Results：The mRNA and protein level of TGF-βandα-SMA increased significantly （P〈0. 01） under the stimulation of high glucose, which was decreased obviously in cordycepin with high glucose group （P〈0. 05）. The expression of E-cadherin mRNA and protein decreased markedly （P〈0. 01） induced by high glucose, which was significantly increased by cordycepinwith high glu-cose group （P〈0. 05）. Conclusion：Cordycepin can significantly inhibit epithelial-mesenchymal-transition of NRK52E cells in-duced by high glucose by downregulate TGF-β expression.