NK1受体及其在P物质和胰岛素样生长因子-1协同增强角膜上皮迁移中的作用。

The NK1 receptor and its participation in the synergistic enhancement of corneal epithelial migration by substance P and insulin-like growth factor-1.

作者信息

Nakamura M, Ofuji K, Chikama T, Nishida T

机构信息

Department of Ophthalmology, Yamaguchi University School of Medicine, Japan.

出版信息

Br J Pharmacol. 1997 Feb;120(4):547-52. doi: 10.1038/sj.bjp.0700923.

DOI:10.1038/sj.bjp.0700923

PMID:9051288

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1564494/

Abstract

We have previously shown that substance P (SP) and insulin-like growth factor-1 (IGF-1) act synergistically to enhance the migration of rabbit corneal epithelial cells in an organ culture model. The present study was designed to identify the epithelial cell SP receptor that participates in this synergistic effect. 2. Rabbit corneal blocks were incubated for 24 h, then the length of the path of epithelial migration was measured. Reagents tried in the TC-199 culture medium, in the presence or absence of IGF-1, were: SP, agonists of tachykinin receptors NK1, NK2 or NK3 and antagonists of tachykinin receptors NK1 or NK2. 3. The binding characteristics of SP receptors were examined in rabbit cultured corneal epithelial cells by binding assays with [125I]-SP in the presence or absence of excess unlabelled SP or ligands of NK1, NK2 or NK3 receptors. 4. As was demonstrated previously, SP and IGF-1 stimulated epithelial migration when they were added to the culture medium together, but individually they had no effect. NK1 agonists had the same synergistic effect with IGF-1 as did SP, but the NK2 and NK3 agonists did not. Furthermore, the NK1 antagonist abolished the synergistic effect of SP and IGF-1, but the NK2 antagonist had no effect. 5. SP bound specifically to rabbit cultured corneal epithelial cells. The binding affinity was 0.44 nM and there were 2.43 x 10(4) binding sites per cell. The NK1 ligand competed, in a dose-dependent fashion, with the binding of SP to corneal epithelial cells, but neither the NK2 nor NK3 ligand affected binding. 6. We conclude that the SP receptor in rabbit corneal epithelial cells is NK1 and that this receptor participates in the synergistic enhancement of corneal epithelial migration by SP and IGF-1. The precise mechanism(s) of this interaction requires more study. These findings imply that both neural and humoral factors are essential for the maintenance and healing of corneal epithelium.

摘要

我们先前已经表明，P物质（SP）和胰岛素样生长因子-1（IGF-1）在器官培养模型中协同作用，增强兔角膜上皮细胞的迁移。本研究旨在鉴定参与这种协同效应的上皮细胞SP受体。2. 将兔角膜块孵育24小时，然后测量上皮迁移路径的长度。在TC-199培养基中，在有或没有IGF-1存在的情况下尝试的试剂有：SP、速激肽受体NK1、NK2或NK3的激动剂以及速激肽受体NK1或NK2的拮抗剂。3. 通过在有或没有过量未标记的SP或NK1、NK2或NK3受体配体存在的情况下用[125I]-SP进行结合试验，在兔培养的角膜上皮细胞中检查SP受体的结合特性。4. 如先前所示，当SP和IGF-1一起添加到培养基中时，它们刺激上皮迁移，但单独添加时它们没有作用。NK1激动剂与SP一样与IGF-1具有相同的协同效应，但NK2和NK3激动剂则没有。此外，NK1拮抗剂消除了SP和IGF-1的协同效应，但NK2拮抗剂没有作用。5. SP特异性结合兔培养的角膜上皮细胞。结合亲和力为0.44 nM，每个细胞有2.43×10(4)个结合位点。NK1配体以剂量依赖性方式与SP与角膜上皮细胞的结合竞争，但NK2和NK3配体均不影响结合。6. 我们得出结论，兔角膜上皮细胞中的SP受体是NK1，并且该受体参与SP和IGF-1对角膜上皮迁移的协同增强作用。这种相互作用的确切机制需要更多的研究。这些发现表明，神经和体液因素对于角膜上皮的维持和愈合都是必不可少的。