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本研究旨在探讨单核细胞趋化蛋白-1(monocyte chemotacitic protein-1,MCP-1)诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,hUVECs)凋亡的分子机制。胶原酶消化收集hUVECs,体外培养细胞,用胰蛋白酶-EDTA混合液消化传代,用血管性假血友病因子(von Willebrand factor,vWF)和VEGF受体2(KDR)免疫染色证实培养细胞为内皮细胞;用不同浓度MCP-1(0.1、1.0、10、100ng/mL)分别作用hUVECs24h、48h;用流式细胞术及蛋白免疫印迹法检测凋亡相关蛋白Fas、Bcl-2、Bax的表达。如我们前期结果所示,MCP-1能诱导hUVECs的凋亡,其效应随浓度和时间的增加而增强;与对照组比较,MCP-1下调抑凋亡蛋白Bcl-2的表达,上调促凋亡蛋白Fas、Bax的表达。以上结果表明,MCP-1能诱导hUVECs凋亡,其作用机制可能与上调Bax、Fas蛋白及下调Bcl-2蛋白表达有关。
This study aimed to investigate the molecular mechanism of monocyte chemotacitic protein-1 (MCP-1) -induced apoptosis in human umbilical vein endothelial cells (hUVECs). HUVECs were collected by collagenase digestion, cultured in vitro, digested and passaged with trypsin-EDTA mixture, and von Willebrand factor (vWF) and VEGF receptor 2 (KDR) immunostaining were used to confirm that cultured cells were endothelium HUVECs were treated with different concentrations of MCP-1 (0.1, 1.0, 10 and 100 ng / mL) for 24 h and 48 h respectively. The expressions of Fas, Bcl-2 and Bax were detected by flow cytometry and Western blotting. As shown in our previous results, MCP-1 induced apoptosis in hUVECs and its effect increased with increasing concentration and time. Compared with the control group, MCP-1 down-regulated the expression of anti-apoptotic protein Bcl-2, Expression of Fas and Bax proteins. The above results showed that MCP-1 can induce apoptosis of hUVECs, and its mechanism may be related to the up-regulation of Bax, Fas protein and down-regulation of Bcl-2 protein expression.