Saito Hajime, Minamiya Yoshihiro, Saito Satoshi, Ogawa Jun-ichi
Second Department of Surgery, Akita University School of Medicine, Akita City, Japan.
J Leukoc Biol. 2002 Oct;72(4):829-36.
The transendothelial migration of neutrophils is a critical step in acute inflammation, which we previously showed to be regulated by endothelial myosin light chain (MLC) kinase. Recent studies suggest that Rho and Rho kinase are also key mediators of MLC phosphorylation, but their roles in neutrophil migration have not been investigated. In the present study, a transwell chamber migration assay system incorporating endothelial monolayer was used to examine the numbers of migrating neutrophils, endothelial F-actin and myosin II rearrangement, and endothelial MLC phosphorylation at selected times during the neutrophil migration in vitro. The results showed that pretreating endothelial cells with C3 (Rho inhibitor) or Y-27632 (Rho kinase inhibitor) significantly diminished neutrophil migration, actin polymerization, myosin II filament formation, and MLC phosphorylation normally associated with the migration. These data suggest that endothelial Rho and Rho kinase regulate transendothelial neutrophil migration by modulating the cytoskeletal events that mediate such migration.
中性粒细胞的跨内皮迁移是急性炎症中的关键步骤，我们之前发现这一过程受内皮肌球蛋白轻链（MLC）激酶调控。近期研究表明，Rho和Rho激酶也是MLC磷酸化的关键介质，但它们在中性粒细胞迁移中的作用尚未得到研究。在本研究中，采用包含内皮单层的Transwell小室迁移分析系统，检测体外中性粒细胞迁移过程中选定时间点的迁移中性粒细胞数量、内皮F-肌动蛋白和肌球蛋白II重排以及内皮MLC磷酸化情况。结果显示，用C3（Rho抑制剂）或Y-27632（Rho激酶抑制剂）预处理内皮细胞，可显著减少中性粒细胞迁移、肌动蛋白聚合、肌球蛋白II丝形成以及通常与迁移相关的MLC磷酸化。这些数据表明，内皮Rho和Rho激酶通过调节介导此类迁移的细胞骨架事件来调控中性粒细胞的跨内皮迁移。
