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目的:本文通过观察血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)影响肾小管上皮细胞转分化(epithelial-mesenchymal transition,EMT)的作用,以及和转化生长因子-β1(transforming growth factor,TGF-β1)作用的关系,探讨AngⅡ参与肾小管间质纤维化的作用机制。方法:以人肾小管上皮细胞株(human kidney cell)HKC细胞为研究对象,采用蛋白印迹等方法,观察AngⅡ(10-9、10-8、10-7、10-6mol/L),及其与TGF-β1共同作用对该细胞表达α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、E-钙黏蛋白(E-cadherin)和纤维连接蛋白(fibronectin,FN)的影响。明胶酶谱法检测细胞培养上清液中基质金属蛋白酶-2和基质金属蛋白酶-9(MMP-2和MMP-9)的变化,Boyden小室检测HKC细胞的迁移能力。结果:①单独应用AngⅡ不能够造成HKC细胞E-cadherin表达的变化,也不能诱导α-SMA表达,但是却能上调FN的表达;②与TGF-β1共同作用时能够加强TGF-β1影响E-cadherin,α-SMA和FN表达的作用;③AngⅡ能够增加HKC生成MMP-2和MMP-9;④AngⅡ能够(10-7和10-6mol/L)增加HKC细胞迁移至Boyden小室膜下侧面的数目。结论:①AngⅡ可以参与EMT过程,但不是导致EMT的关键因素;②AngⅡ能够以协同的方式参与TGF-β1导致的EMT,可能以此方式加重肾小管间质的纤维化。
Objective: To observe the effect of angiotensin Ⅱ (AngⅡ) on the epithelial-mesenchymal transition (EMT) and the effect of transforming growth factor-β1 (TGF-β1) The relationship between Ang Ⅱ and tubulointerstitial fibrosis was investigated. Methods: Human renal cell HKC cells were used as research objects. AngⅡ (10-9,10-8,10-7,10-6 mol / L) and On the expression of α-smooth muscle actin (α-SMA), E-cadherin and fibronectin (FN) with TGF-β1. Gelatin zymography was used to detect the changes of matrix metalloproteinase-2 and matrix metalloproteinase-9 (MMP-2 and MMP-9) in cell culture supernatants. Boyden chamber was used to detect the migration of HKC cells. Results: ① AngⅡ alone could not change the expression of E-cadherin, induce the expression of α-SMA, but up-regulate the expression of FN in HKC cells; ② When combined with TGF-β1, the effect of TGF- cadherin, α-SMA and FN; ③ AngⅡ increased the production of MMP-2 and MMP-9 by HKC; ④ AngⅡ could increase the number of HKC cells migrating to the inferior side of Boyden’s subventricular zone (10-7 and 10-6 mol / L) Conclusion AngⅡ can participate in EMT but not the key factor leading to EMT. AngⅡ can participate in TGF-β1-induced EMT in a synergistic manner, which may aggravate tubulointerstitial fibrosis in this way.