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目的探讨粉防己碱(tetrandrine,Tet)对转化生长因子-β1(TGF-β1)诱导的人近端肾小管上皮细胞(HK-2)转分化的拮抗效应及其机制。方法体外培养的HK-2随机分为正常对照组、Tet组(100ng/ml)、TGF-β1(5ng/ml)组和TGF-β1(5ng/ml)+Tet(100ng/ml)组。Westernblot法检测细胞中TβRI蛋白、SnoN蛋白水平、Smad7蛋白水平和Smad2/3磷酸化水平的改变;半定量反转录-聚合酶链式反应(RT-PCR)方法观察细胞中TβRI mRNA、Smad7 mRNA、SnoN mRNA表达的改变。免疫荧光检测间充质细胞标记物α-平滑肌肌动蛋白(α-SMA)的表达。结果 Tet能抑制TGF-β1诱导α-SMA的表达,显著上调SnoN mRNA和蛋白的表达(P<0.01);Tet对TβRI表达、Smad2/3的磷酸化及Smad7的表达没有影响。结论 Tet能抑制人近端小管上皮细胞转分化,其机制与Tet上调SnoN表达有关。
Objective To investigate the antagonistic effect of tetrandrine (Tet) on transdifferentiation of human proximal tubular epithelial cells (HK-2) induced by transforming growth factor-β1 (TGF-β1) and its mechanism. Methods HK-2 cultured in vitro was randomly divided into normal control group, Tet group (100ng / ml), TGF-β1 (5ng / ml) group and TGF-β1 (5ng / ml) The levels of TβRI protein, SnoN protein, Smad7 protein and Smad2 / 3 phosphorylation were detected by Western blotting. The expressions of TβRI mRNA and Smad7 mRNA were detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) , SnoN mRNA expression changes. Immunofluorescence was used to detect the expression of mesenchymal cell marker α-smooth muscle actin (α-SMA). Results Tet could inhibit the expression of α-SMA induced by TGF-β1 and up-regulate the expression of SnoN mRNA and protein (P <0.01). Tet did not affect the expression of TβRI, phosphorylation of Smad2 / 3 and Smad7. Conclusion Tet can inhibit the transdifferentiation of human proximal tubular epithelial cells, and its mechanism is related to Tet up-regulation of SnoN expression.