Release of pro- and anti-angiogenic factors by human cardiac fibroblasts: effects on DNA synthesis and protection under hypoxia in human endothelial cells.
Release of pro- and anti-angiogenic factors by human cardiac fibroblasts: effects on DNA synthesis and protection under hypoxia in human endothelial cells.
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DOI：
10.1016/s0167-4889(01)00078-7
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发表时间：
2001-04
期刊：
影响因子：
--
通讯作者：
L. Zhao;M. Eghbali-Webb
中科院分区：
文献类型：
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作者：
L. Zhao;M. Eghbali-Webb
心肌由不同的细胞类型组成，提示细胞间的相互作用在维持心脏的结构和功能完整性方面起着重要作用。心脏成纤维细胞是心脏中细胞外基质、生长因子和细胞因子的来源，并认识到它们与心肌细胞的相互作用。然而，它们对血管内皮细胞生物学反应的影响还远未被探索。血管内皮细胞的增殖是血管生成的重要阶段，也是冠脉侧支发育的重要环节。本研究旨在探讨心脏成纤维细胞产生的可溶性因子对内皮细胞增殖的影响。人心脏成纤维细胞条件培养液(CF-CM)可使人脐静脉内皮细胞(HUVECDNA)合成增加47%(P<0.0001)。这一作用依赖于从头蛋白的合成和MAP激酶的激活。CF-CM可诱导HUVEC中ERK2的表达和活化。去掉肝素结合蛋白的CF-CM对人脐静脉内皮细胞DNA合成的刺激作用明显高于“整体CF-CM”。免疫印迹分析显示整个CFCM中有血管内皮生长因子、碱性成纤维细胞生长因子、血小板衍生生长因子、转化生长因子-β-1、纤维连接蛋白和凝血酶反应蛋白-1的表达。对整个CF-CM中各个因子的免疫耗竭实验表明，所有因子都是CF-CM充分活性所必需的。CF-CM可显著逆转缺氧对HUVEC DNA合成的抑制作用，增强细胞存活相关蛋白Bcl2的表达。总之，这些数据表明，心脏成纤维细胞释放抑制和刺激因子，其净效应是促进内皮细胞的DNA合成。这些结果表明心脏成纤维细胞可能在心脏血管生成中发挥作用。
Myocardium consists of diverse cell types suggesting a role for cell-cell interaction in maintaining the structural and functional integrity of the heart. Cardiac fibroblasts are the source of extracellular matrix, growth factors and cytokines in the heart and their interactions with cardiac myocytes are recognized. Their effects on biological responses of endothelial cells, however, are vastly unexplored. Proliferation of endothelial cells is an essential stage of angiogenesis and contributes to development of coronary collaterals. This study was designed to evaluate the effect of soluble factors produced by cardiac fibroblasts on endothelial cell proliferation. Human cardiac fibroblast-conditioned medium (CF-CM) caused a significant increase (47%, P<0.0001) in DNA synthesis in human umbilical vein endothelial cells (HUVEC), as determined by [3H]thymidine incorporation. This effect was dependent on de novo protein synthesis and activation of MAP kinases. Consistently, CF-CM induced the expression and activation of ERK2 in HUVEC. The CF-CM from which heparin-binding proteins were removed, had a significantly enhanced stimulatory effect on DNA synthesis in HUVEC compared to that of ‘whole CF-CM’. Western analysis showed the presence of VEGF, bFGF, PDGF, TGF-β1, fibronectin and thrombospondin-1 in whole CF-CM. The individual immunodepletion of each factor from whole CF-CM showed that all were necessary for full activity of CF-CM. CF-CM caused a significant reversal of hypoxia-induced inhibition of DNA synthesis and enhanced expression of survival-associated protein, Bcl2, in HUVEC. Together, these data show that cardiac fibroblasts release inhibitory and stimulatory factors, the net effect of which is an enhancement of DNA synthesis in endothelial cells. These results point to the role that cardiac fibroblasts may play in angiogenesis in the heart.