Thrombin-induced gap formation in confluent endothelial cell monolayers in vitro.
Thrombin-induced gap formation in confluent endothelial cell monolayers in vitro.
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DOI：
10.1182/blood.v62.3.549.549
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发表时间：
1983-09
期刊：
影响因子：
20.3
通讯作者：
M. Laposata;D. K. Dovnarsky;H. Shin
中科院分区：
文献类型：
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作者：
M. Laposata;D. K. Dovnarsky;H. Shin
当凝血酶在体外与人脐静脉内皮细胞的汇合单层孵育时，内皮细胞的形状发生变化，导致单层中出现间隙，破坏内皮的完整性并暴露内皮下。使用网格测定来测量这种现象，我们观察到，在与10（-2）U/ml（10（-10）M）凝血酶孵育15分钟后，高达80%的表面积一旦被细胞覆盖就被揭开。该效果在2分钟内明显，并且没有从培养皿表面去除细胞。在暴露于凝血酶后2小时内，单层中的间隙完全消失。凝血酶的作用被抑制凝血酶与水蛭素或抗凝血酶III加肝素或预孵育的单细胞层与二丁酰环磷酸腺苷（dbcAMP）。组胺也诱导内皮细胞单层间隙形成。吡拉明和西咪替丁都阻止组胺诱导的作用，但它们对凝血酶诱导的间隙形成没有影响。完整的单层细胞未被缓激肽、5-羟色胺、C5 a或C3 a破坏。我们的研究结果表明，少量的凝血酶可以诱导反复和短暂暴露的内皮下，这种情况被认为是有利于动脉粥样硬化和血栓形成。
When thrombin is incubated with confluent monolayers of human umbilical vein endothelial cells in vitro, there is a change in the shape of the endothelial cells that results in gaps in the monolayer, disrupting the integrity of the endothelium and exposing the subendothelium. Using a grid assay to measure this phenomenon, we observed that up to 80% of the surface area once covered by cells was uncovered after a 15-min incubation with 10(-2) U/ml (10(-10)M) thrombin. The effect was apparent within 2 min and did not remove cells from the surface of the culture dish. The gaps in the monolayer completely disappeared within 2 hr after exposure to thrombin. The effect of thrombin was inhibited by preincubation of thrombin with hirudin or antithrombin III plus heparin or by preincubation of the monolayers with dibutyryl cyclic adenosine monophosphate (dbcAMP). Histamine also induced gap formation in endothelial cell monolayers. Both pyrilamine and cimetidine prevented the histamine-induced effect, but they had no effect on thrombin-induced gap formation. Intact monolayers were not disrupted by bradykinin, serotonin, C5a, or C3a. Our results suggest that small amounts of thrombin can induce repeated and transient exposure of the subendothelium, a situation believed to be conducive to atherogenesis and thrombosis.