Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity.
Cooperative effect of TNFalpha, bFGF, and VEGF on the formation of tubular structures of human microvascular endothelial cells in a fibrin matrix. Role of urokinase activity.
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DOI：
10.1083/jcb.132.6.1177
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发表时间：
1996-03
期刊：
影响因子：
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通讯作者：
van Hinsbergh VW
中科院分区：
文献类型：
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作者：
Koolwijk P;van Erck MG;de Vree WJ;Vermeer MA;Weich HA;Hanemaaijer R;van Hinsbergh VW
在与组织修复和疾病相关的血管生成中，纤维蛋白和炎症介质经常参与。我们使用三维纤维蛋白基质研究了人微血管内皮细胞（hMVEC）形成毛细血管样管状结构所需的体液。bFGF和VEGF165不能自行诱导出管状结构。其中一种或两种因子与TNFalpha同时加入可诱导小管生长，其中bFGF、VEGF165和TNFalpha同时加入效果最强。外源添加的u-PA，而不是其非蛋白水解的氨基末端片段，可以取代TNFalpha，这表明TNFalpha诱导的u-PA合成参与其中。可溶性u- pa受体（u- PAR）或抑制u- pa活性的抗体可阻止59-99%的管状结构的形成。epsilon-ACA和trasylol抑制了纤溶酶的形成和活性，使管的形成程度降低了71-95%。如果没有额外的生长因子，TNFalpha或u-PA不会诱导管状结构。bFGF和VEGF165分别使u-PAR增加72%和46%，而TNFalpha本身也使hMVEC的u-PAR增加30%。诱导有丝分裂并不是bFGF和VEGF165的主要贡献，因为细胞数量在TNFalpha存在下没有明显变化，而完全抑制有丝分裂的tyrphostin A47仅减少了28-36%的管状结构的形成。这些数据表明，除了血管生成因子VEGF165和/或bFGF外，细胞因子TNFalpha还需要诱导细胞结合的u-PA活性，以诱导hMVEC在纤维蛋白基质中体外形成毛细血管样结构。这些数据可能为在病理条件下血管生成的机制提供见解。
In angiogenesis associated with tissue repair and disease, fibrin and inflammatory mediators are often involved. We have used three- dimensional fibrin matrices to investigate the humoral requirements of human microvascular endothelial cells (hMVEC) to form capillary-like tubular structures. bFGF and VEGF165 were unable to induce tubular structures by themselves. Simultaneous addition of one or both of these factors with TNFalpha induced outgrowth of tubules, the effect being the strongest when bFGF, VEGF165, and TNFalpha were added simultaneously. Exogenously added u-PA, but not its nonproteolytic amino-terminal fragment, could replace TNFalpha, suggesting that TNFalpha-induced u-PA synthesis was involved. Soluble u-PA receptor (u- PAR) or antibodies that inhibited u-PA activity prevented the formation of tubular structures by 59-99%. epsilon-ACA and trasylol which inhibit the formation and activity of plasmin reduced the extent of tube formation by 71-95%. TNFalpha or u-PA did not induce tubular structures without additional growth factors. bFGF and VEGF165 enhanced of the u- PAR by 72 and 46%, but TNFalpha itself also increased u-PAR in hMVEC by 30%. Induction of mitogenesis was not the major contribution of bFGF and VEGF165 because the cell number did not change significantly in the presence of TNFalpha, and tyrphostin A47, which inhibited mitosis completely, reduced the formation of tubular structures only by 28-36%. These data show that induction of cell-bound u-PA activity by the cytokine TNFalpha is required in addition to the angiogenic factors VEGF165 and/or bFGF to induce in vitro formation of capillary-like structures by hMVEC in fibrin matrices. These data may provide insight in the mechanism of angiogenesis as occurs in pathological conditions.