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(英文)  

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

作　　者：李琴山[1] 刘洋[1] 冯赞杰[1] 卢志顺[1] 钱民章[1] 

机构地区：[1]遵义医学院生物化学与分子生物学教研室,遵义563003

出　　处：《生理学报》2010年第1期63-68,共6页Acta Physiologica Sinica

基　　金：supported by the National Natural Science Foundation of China(No.30760078)

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 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。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 identified by 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 by flow cytometry （FACS） and Western blot.As shown in our preliminary study,MCP-1 induced apoptosis of hUVECs in a dosedependent manner at both 24 h and 48 h.FACS and Western blot analysis results in the present study indicated that MCP-1 promoted the expression of proapoptotic proteins Bax and Fas and inhibited the expression of antiapoptotic protein Bcl-2.These results suggest that MCP-1 may induce the apoptosis of hUVECs through evoking the imbalance between proapoptotic Fas/Bax and antiapoptotic Bcl-2 protein.

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[医药卫生—人体解剖和组织胚胎学]