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.
Sheng Li Xue Bao. 2010 Feb 25;62(1):63-8.
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 dose-dependent 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.
本研究旨在探讨Bcl-2、Fas和Bax是否参与单核细胞趋化蛋白-1（MCP-1）诱导的人脐静脉内皮细胞（hUVECs）凋亡。培养hUVECs，并用特异性抗血管性血友病因子（vWF）和抗血管内皮生长因子受体-2（KDR）抗体通过免疫荧光和免疫组织化学鉴定其纯度。当hUVECs汇合度达到90%时，使其血清饥饿12小时，然后分别用不同浓度的MCP-1（0.1、1.0、10、100 ng/mL）处理24小时和48小时。通过流式细胞术（FACS）和蛋白质免疫印迹法检测凋亡相关蛋白Fas、Bcl-2、Bax的表达。如我们的初步研究所示，MCP-1在24小时和48小时均以剂量依赖性方式诱导hUVECs凋亡。本研究中的FACS和蛋白质免疫印迹分析结果表明，MCP-1促进促凋亡蛋白Bax和Fas的表达，并抑制抗凋亡蛋白Bcl-2的表达。这些结果表明，MCP-1可能通过引发促凋亡的Fas/Bax和抗凋亡的Bcl-2蛋白之间的失衡来诱导hUVECs凋亡。
