Migration of eosinophils through basement membrane components in vitro: Role of matrix metalloproteinase-9
Migration of eosinophils through basement membrane components in vitro: Role of matrix metalloproteinase-9
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DOI：
10.1165/ajrcmb.17.4.2877
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发表时间：
1997-10-01
影响因子：
6.4
通讯作者：
Leiferman, KM
中科院分区：
文献类型：
--
作者：
Okada, S;Kita, H;Leiferman, KM
一般来说，炎症细胞通过产生蛋白酶而穿过基底膜。为了研究蛋白酶在嗜酸性粒细胞基底膜迁移中的作用，我们在Matrigel（(R)）包被的趋化室中研究了外周血嗜酸性粒细胞。电镜显示，当添加到上腔的嗜酸性粒细胞在下腔的血小板活化因子（PAF）和两个腔的白细胞介素(IL)-5存在的情况下迁移膜时，基质层(R)层降解。在没有PAF和IL-5或两者都没有的情况下，Matrigel(R)层没有发生变化。PAF和IL-5诱导的嗜酸性粒细胞的迁移被1,10-菲罗啉、batimastat、3,4-二氯异acoumarin、chymostatin和基质金属蛋白酶(MMP)-9的中和抗体抑制，表明丝氨酸蛋白酶(s)和MMP，特别是MMP-9，参与了通过Matrigel（(R)）介导的嗜酸性粒细胞的迁移。相反，嗜酸性粒细胞通过裸膜的迁移不受batimastat的影响。采用明胶酶谱法和免疫印迹法，在嗜酸性粒细胞迁移实验的上腔上清液和嗜酸性粒细胞条件培养基中检测到MMP-9。IL-5、PAF或两者均能增加MMP-9的释放，但只有IL-5和PAF同时存在时，MMP-9的底物降解活性才会增加，这表明MMP-9的释放和激活机制参与了嗜酸性细胞基底膜迁移。本研究提示MMP-9参与嗜酸性粒细胞的基底膜迁移，并提示其参与组织嗜酸性粒细胞发挥作用的炎症性疾病。
In general, inflammatory cells cross basement membranes by producing proteinases. To investigate the role of proteinases in eosinophil basement membrane migration, we studied peripheral blood eosinophils in Matrigel((R))-coated chemotaxis chambers. Electron microscopy showed degradation of the Matrigel((R)) layer when eosinophils, added to the upper chamber, transmigrated the membrane in the presence of both platelet-activating factor (PAF) in the lower chamber and interleukin (IL)-5 in both chambers. In the absence of either or both PAF and IL-5, no changes occurred in the Matrigel((R)) layer. Matrigel((R)) transmigration of eosinophils induced by PAF and IL-5 was inhibited by 1,10-phenanthroline, batimastat, 3,4-dichloroisacoumarin, chymostatin, and a neutralizing antibody for the matrix metalloproteinase (MMP)-9, indicating that serine proteinase(s) and MMP, specifically MMP-9, were involved in the transmigration of eosinophils through Matrigel((R)). In contrast, eosinophil migration through a bare membrane was not affected by batimastat. Using gelatin zymography and immunoblotting, MMP-9 was detected in the migration upper chamber supernatant of the eosinophil transmigration assay and in the conditioned medium of eosinophils. Release of MMP-9 by eosinophils was increased by IL-5, PAF, or both, but the substrate-degrading activity of MMP-9 was increased only in the presence of both IL-5 and PAF, indicating that the releasing and activating mechanisms of MMP-9 are involved in eosinophil basement membrane migration. This study implicates MMP-9 in basement membrane migration of eosinophils and suggests its involvement in inflammatory diseases where tissue eosinophilia plays a role.