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单核细胞趋化蛋白-1 诱导人脐静脉内皮细胞凋亡的分子机制

李琴山, 刘洋, 冯赞杰, 卢志顺, 钱民章

遵义医学院生物化学与分子生物学教研室，遵义 563003

摘要

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 umbilicalvein 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、100 ng/mL)分别作用hUVECs 24 h、48 h；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。

关键词： 单核细胞趋化蛋白-1; 人脐静脉内皮细胞; 动脉粥样硬化; Bax 蛋白; Fas 蛋白; Bcl-2 蛋白; 凋亡

分类号：R329.2

The molecular mechanism of apoptosis of human umbilical veinendothelial cells induced by monocyte chemotacitic protein-1

LI Qin-Shan, LIU Yang, FENG Zan-Jie, LU Zhi-Shun, QIAN Min-Zhang

Department of Biochemistry and Molecular Biology, Zunyi Medical College, Zunyi 563003, China

Abstract

The present study was aimed to investigate whether Bcl-2, Fas and Bax are involved in monocyte chemotacitic protein-1(MCP-1)-induced apoptosis of human umbilical vein endothelial cells (hUVECs). hUVECs were cultured, and the purity was identifiedby immunofluorescence and immunohistochemistry with specific anti-von Willebrand factor (vWF) and anti-VEGF receptor-2 (KDR)antibodies. With 90% confluence hUVECs were serum-starved for 12 h, and then treated with different concentrations of MCP-1(0.1, 1.0, 10, 100 ng/mL) for 24 and 48 h respectively. The expressions of apoptosis related proteins Fas, Bcl-2, Bax were detected byflow cytometry (FACS) and Western blot. As shown in our preliminary study, MCP-1 induced apoptosis of hUVECs in a dosedependentmanner at both 24 h and 48 h. FACS and Western blot analysis results in the present study indicated that MCP-1 promotedthe expression of proapoptotic proteins Bax and Fas and inhibited the expression of antiapoptotic protein Bcl-2. These results suggestthat MCP-1 may induce the apoptosis of hUVECs through evoking the imbalance between proapoptotic Fas/Bax and antiapoptoticBcl-2 protein.

Key words: monocyte chemotacitic protein-1; human umbilical vein endothelial cell; atherosclerosis; Bcl-2; Bax; Fas; apoptosis

收稿日期：2009-09-03　　录用日期：2010-01-04

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引用本文：

李琴山, 刘洋, 冯赞杰, 卢志顺, 钱民章. 单核细胞趋化蛋白-1 诱导人脐静脉内皮细胞凋亡的分子机制[J]. 生理学报 2010; 62 (1): 63-68.

LI Qin-Shan, LIU Yang, FENG Zan-Jie, LU Zhi-Shun, QIAN Min-Zhang. The molecular mechanism of apoptosis of human umbilical veinendothelial cells induced by monocyte chemotacitic protein-1. Acta Physiol Sin 2010; 62 (1): 63-68 (in Chinese with English abstract).