【摘要】 [目的]研究三七皂苷（Panax notoginseng saponins,PNS）对高糖诱导后肾小管上皮细胞发生上皮细胞-间充质转化（epithelial-mesenchymal transition,EMT）的修复作用及其相关机制。[方法]培养肾小管上皮细胞株NRK-52E细胞,分为正常对照组、高糖组、白藜芦醇（resveratrol,RSV）组、PNS组、沉默信息调节因子1（silent information regulator 1,SIRT1）抑制剂EX527组和EX527+PNS组。药物干预48h后收集细胞;采用免疫组化检测α-平滑肌肌动蛋白（alpha-smooth muscle actin,α-SMA）、E-钙黏着蛋白（E-cadherin,E-cad）的蛋白表达;Real-time RT-PCR检测SIRT1、α-SMA、E-cad、转化生长因子-β₁（transforming growth factor-β₁,TGF-β₁）、Smad3、Smad4的基因表达,Western blot检测SIRT1、α-SMA、E-cad、TGF-β₁的蛋白表达。[结果]与正常对照组比较,高糖诱导后的NRK-52E细胞出现明显的EMT表现,即α-SMA蛋白表达增多（P<0.05）,E-cad蛋白表达减少（P<0.01）。与高糖组比较,PNS组α-SMA蛋白表达减少（P<0.05）,E-cad和SIRT1蛋白表达增加（P<0.01,P<0.01）,TGF-β₁、Smad3、Smad4基因表达降低（P<0.01,P<0.05）。EX527干预后,PNS作用受到抑制。[结论]PNS对肾小管上皮细胞具有保护作用,其机制可能是通过激活SIRT1抑制TGF-β₁/Smad通路,从而阻止高糖诱导的EMT的发生。更多还原

【Abstract】 [Objective] To investigate the repair effect of the panax notoginseng saponins（PNS） on epithelial-mesenchymal transition of rat renal tubular epithelial cells（NRK-52 E） induced by high glucose and from the SIRT1/TGF-β₁/Smad signaling pathway. [Methods]NRK-52 E were cultured in DMEM medium with 10% fetal bovine serum, and divided into normal control group, high glucose group, resveratrol（RSV） group, PNS group, EX527 group and EX527 + PNS group and collected cells after 48 hours of drug intervention. The expression of the α-SMA and E-cadherin（E-cad） was also examined by immunohistochemistry. In addition, SIRT1, α-SMA, E-cad, TGF-β₁, Smad3 and Smad4 gene expression was analyzed by Real-time RT-PCR. The protein expressions of SIRT1, α-SMA, E-cad, TGF-β₁ were determined by Western blot. [Results] Compared with normal control group, the high glucose induced cells appeared α-SMA increase（P<0.05） and E-cad decrease（P<0.01）. Compared with high glucose group, PNS intervention resulted in the decrease of α-SMA（P<0.05）,the increase of E-cad（P<0.01）, the increases of SIRT1（P<0.01）, the decreases of TGF-β₁, Smad3 and Smad4（P<0.01, P<0.05）. However,PNS had not obvious effect on the cells under the intervention of the SIRT1 blocker EX527. [Conclusion] PNS has a protective effect on the epithelial cells.The mechanism may be through the activation of SIRT1 and inhibition of TGF-β₁/Smad pathway, thereby to prevent the high glucose-induced EMT of epithelial cells.更多还原