Noiri E, Hu Y, Bahou W F, Keese C R, Giaever I, Goligorsky M S
Department of Medicine, State University of New York, Stony Brook, New York 11794-8152, USA.
J Biol Chem. 1997 Jan 17;272(3):1747-52. doi: 10.1074/jbc.272.3.1747.
Endothelin (ET) synthesis is enhanced at sites of ischemia or in injured vessels. The purpose of this study was to explore the possibility of autocrine stimulation of endothelial cell migration by members of the endothelin family. Experiments with microvascular endothelial cell transmigration in a Boyden chemotactic apparatus showed that endothelins 1 and 3, as well as a selective agonist of ETB receptor IRL-1620, equipotently stimulated migration. Endothelial cell migration was unaffected by the blockade of ETA receptor, but it was inhibited by ETB receptor antagonism. Based on our previous demonstration of signaling from the occupied ETB receptor to constitutive nitric oxide (NO) synthase (Tsukahara, H., Ende, H., Magazine, H. I., Bahou, W. F., and Goligorsky, M. S. (1994) J. Biol. Chem. 269, 21778-21785), we next examined the contribution of ET-stimulated NO production to endothelial cell migration. In three independent cellular systems, 1) migration and wound healing by microvascular endothelial cells, 2) wound healing by Chinese hamster ovary cells stably expressing ETB receptor with or without endothelial NO synthase, and 3) application of antisense oligodeoxynucleotides targeting endothelial NO synthase in human umbilical vein endothelial cells, an absolute requirement for the functional NO synthase in cell migration has been demonstrated. These findings establish the permissive role of NO synthesis in endothelin-stimulated migration of endothelial cells.
内皮素（ET）的合成在缺血部位或受损血管中会增强。本研究的目的是探讨内皮素家族成员自分泌刺激内皮细胞迁移的可能性。在Boyden趋化装置中进行的微血管内皮细胞迁移实验表明，内皮素-1和内皮素-3以及ETB受体的选择性激动剂IRL-1620具有同等效力地刺激迁移。内皮细胞迁移不受ETA受体阻断的影响，但受ETB受体拮抗作用的抑制。基于我们之前关于被占据的ETB受体向组成型一氧化氮（NO）合酶发出信号的证明（Tsukahara, H., Ende, H., Magazine, H. I., Bahou, W. F., and Goligorsky, M. S. (1994) J. Biol. Chem. 269, 21778 - 21785），接下来我们研究了ET刺激产生的NO对内皮细胞迁移的作用。在三个独立的细胞系统中，1）微血管内皮细胞的迁移和伤口愈合，2）稳定表达ETB受体且有或没有内皮型NO合酶的中国仓鼠卵巢细胞的伤口愈合，以及3）在人脐静脉内皮细胞中应用靶向内皮型NO合酶的反义寡脱氧核苷酸，已证明细胞迁移对功能性NO合酶有绝对需求。这些发现确立了NO合成在内皮素刺激的内皮细胞迁移中的许可作用。
