Knock-down of endothelial connexins impairs angiogenesis
Knock-down of endothelial connexins impairs angiogenesis
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DOI：
10.1016/j.phrs.2011.11.012
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发表时间：
2012-03-01
影响因子：
9.3
通讯作者：
Dhein, Stefan
中科院分区：
文献类型：
--
作者：
Gaertner, Christiane;Ziegelhoeffer, Barbara;Dhein, Stefan
连接蛋白(Cx)被认为在生长和分化中起重要作用。通过人脐静脉内皮细胞(HUVEC)18h Matrigel体外血管生成实验，探讨内皮Cx在血管生成过程中的作用。在Matrigel细胞上培养之前，用尼古丁或缝隙连接抑制剂棕榈油酸(PA)或Cx37、Cx40或Cx43的siRNA下调。免疫荧光显微镜观察Cx表达的变化及其对缝隙连接通讯的影响。Western印迹和荧光黄转移。敲除每一种Cx-亚型均可显著降低HUVEC中Cx蛋白的含量。Cx-基因敲除和PA处理均可损害通过缝隙连接的细胞间通讯，并显著减少毛细血管分支数量。下调Cx43和Cx40或用PA处理降低了血管生成分析中的复杂模式，尼古丁显著减少了Cx43和Cx37的表达，并显著减少了Matrigel实验中的平均毛细血管长度、分支数量和模式。我们可以得出结论，连接蛋白参与了血管生成，特别是分支的形成。这可以部分解释尼古丁治疗下血管生成的变化。(C)2011爱思唯尔有限公司。保留所有权利。
Connexins (Cx) are suggested to play important roles in growth and differentiation. Aim of our study was to investigate the role of endothelial Cx in the angiogenic process.Several parameters of angiogenesis were assessed in 18 h Matrigel in vitro angiogenesis assays with human umbilical vein endothelial cells (HUVEC). Prior to culture on Matrigel cells were treated with nicotine or the gap junction inhibitor palmitoleic acid (PA), or siRNA-knock-down of either Cx37, Cx40 or Cx43 was performed. Changes in Cx expression and their effects on gap-junctional communication were investigated using immunofluorescence microscopy. Western blot and Lucifer Yellow dye transfer.Knock-down of each Cx-isoform significantly reduced the amount of specific Cx protein in HUVEC. Cx-knock-down as well as treatment with PA impaired intercellular communication via gap junctions and diminished significantly the number of capillary branches. Knock-down of Cx43 and Cx40 or treatment with PA reduced complexity pattern in the angiogenesis assay.Nicotine significantly reduced expression of Cx43 and Cx37 as well as average length of capillary branches, number of branches and pattern in the Matrigel assay. We can conclude that connexins are involved in angiogenesis, in particular in branch formation. This can partly explain the changes in angiogenesis seen under nicotine treatment. (C) 2011 Elsevier Ltd. All rights reserved.