HGF上调SnoN mRNA抑制高糖介导大鼠肾小管上皮细胞纤维病变机制

Study on the Mechanism of HGF Upregulate SnoN mRNA to Inhibit High Glucose Induced Rat Renal Tubular Epithelial Cells Fibrosis

目的:探讨肝细胞生长因子(HGF)是否通过转录共抑制因子(SnoN)影响高糖介导大鼠肾小管上皮细胞纤维病变。方法:将体外培养NRK-52E细胞分为对照组(NG组,正常糖培养液培养)、高糖对照组(HG组,高糖培养液培养)、10μg/LrhHGF干预组(低剂量rhHGF组,10μg/LrhHGF高糖培养液培养)及20μg/LrhHGF干预组(高剂量rhHGF组,20μg/LrhHGF高糖培养液培养);培养48h时,采用Westernblot方法检测各组细胞SnoN、细胞外信号调节激酶1和2(ERK1/2)信号通路及E-钙黏素(E-cadherin)、α-平滑肌肌动蛋白(α-SMA)及胶原蛋白Ⅲ(Col-Ⅲ)蛋白的表达,qRT-PCR技术检测SnoNmRNA的表达。结果:与NG组相较,HG组细胞中α-SMA、Col-Ⅲ蛋白和活化的ERK1/2表达上调(P<0.05),E-cadherin、SnoN蛋白表达下调(P<0.05),SnoNmRNA表达增高(P<0.05);然而与HG组比较,在rhHGF干预组,10μg/L或20μg/LrhHGF均可使NRK-52E细胞α-SMA和Col-Ⅲ蛋白表达减少(P<0.05),而E-cadherin、SnoN蛋白和活化的ERK1/2表达增多(P<0.05),SnoNmRNA表达进一步增高(P<0.05)。结论:HGF阻断或对抗高糖介导肾小管上皮细胞-肌成纤维细胞转分化和间质细胞外基质的沉积,其机制可能是通过活化ERK1/2通路上调SnoNmRNA表达实现。

Objective:To investigate whether hepatocyte growth factor(HGF)affect rat renal tubular epithelial fibrosis through transcriptional co-inhibitor SnoN(Ski-related novel protein N).Methods:NRK-52E cells were cultured in vitro were divided into control group(NG group,cultivated in normal sugar solution),high sugar group(HG group,cultivated in high sugar solution),10μg/L rhHGF intervening group(low dose rhHGF group,cultivated in 10μg/L rhHGF high dose sugar solution)and 20μg/L rhHGF intervening group(high dose rhHGF group,cultivated in 20μg/L rhHGF high dose sugar solution).After cultivated for 48 h,Western blot was used to detect SnoN,extracellular signal-regulated kinase-1 and-2(ERK1/2)signaling pathway,E-cadherin andα-smooth muscle actin(α-SMA)and Collagen III(Col-III)protein expression;qRT-PCR technique was used to detect the expression of SnoN mRNA.Results:Compared with NG group,the expression ofα-SMA,Col-Ⅲand activated ERK1/2 was up-regulated in HG group(P<0.05),while the expression of E-cadherin and SnoN protein was down-regulated(P<0.05);the expression of SnoN mRNA was increased(P<0.05).However,compared with HG group,in rhHGF group,10μg/L or 20μg/L rhHGF could lower the expression ofα-SMA and Col-Ⅲprotein(P<0 05),while the expression of E-cadherin,SnoN and activated ERK1/2 increased(P<0.05);SnoN mRNA expression was further improved(P<0.05).Conclusion:HGF blocks or antagonizes the process of epithelial-mesenchymal transition(EMT)and the deposition of interstitial extracellular matrix(ECM)in renal tubular epithelial cells mediated by high glucose by activating ERK1/2 pathway to up-regulate the expression of SnoNmRNA.