| Rho激酶信号通路参与ET-1诱导的人气道平滑肌细胞迁移和细胞骨架的变化 |
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| 引用本文： | 黎振兴,罗雅玲,赖文岩,徐健,任敦强,赵燕霞. Rho激酶信号通路参与ET-1诱导的人气道平滑肌细胞迁移和细胞骨架的变化[J]. 南方医科大学学报, 2008, 28(6): 1031-1034 |
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| 作者姓名： | 黎振兴 罗雅玲 赖文岩 徐健 任敦强 赵燕霞 |
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| 作者单位： | 南方医科大学南方医院呼吸内科,广东,广州,510515;南方医科大学南方医院心血管内科重点实验室,广东,广州,510515;南方医科大学南方医院肾移植科,广东,广州,510515 |
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| 摘 要： | 目的 研究Rho激酶信号通路在内皮素-1(ET-1)诱导的人气道平滑肌细胞(ASMCs)迁移及细胞骨架变化中的作用.方法 组织块贴壁法培养人ASMCs,改良的Boyden小室检测ASMCs的迁移能力;激光共聚焦扫描显微镜观察细胞骨架的变化;Western blotting检测ET-1刺激不同时间后Rho激酶底物肌球蛋白磷酸酶目标亚单位1(MYPT1)的磷酸化水平.结果 ET-1在0.1、1、10、100 nmol/L浓度具有趋化ASMCs的迁移能力,10 nmol/L的ET-1趋化ASMCs迁移的能力最强(与对照组相比,P<0.01).Rho激酶抑制剂Y-27632可浓度依赖性地抑制ET-1趋化的ASMCs跨膜迁移,与ET-1组相比,10μmol/L的Y-27632显著抑制了ASMCs迁移(P<0.01);ET-1(10 nmol/L)刺激后ASMCs细胞骨架和形态改变,伪足生成,应力纤维形成增多,Y-27632可显著抑制ET-1诱导的上述改变;ET-1刺激15、30min后p-MYPT1表达显著增高(与对照组相比,P<0.01),随着刺激时间的延长,p-MYPT1表达下降(P>05).结论 Rho激酶信号通路在ET-1诱导的ASMCs迁移和细胞骨架变化中发挥重要的作用.
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| 关 键 词： | 人气道平滑肌细胞 内皮素-1 迁移 细胞骨架 Rho激酶 |
| 文章编号： | 1673-4254(2008)06-1031-04 |
| 修稿时间： | 2008-04-07 |
Rho-kinase signaling pathway participates in endothelin-1-induced human airway smooth muscle cell migration and cytoskeletal reorganization |
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| LI Zhen-xing,LUO Ya-ling,LAI Wen-yan,XU Jian,REN Dun-qiang,ZHAO Yan-xia. Rho-kinase signaling pathway participates in endothelin-1-induced human airway smooth muscle cell migration and cytoskeletal reorganization[J]. Journal of Southern Medical University, 2008, 28(6): 1031-1034 |
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| Authors: | LI Zhen-xing LUO Ya-ling LAI Wen-yan XU Jian REN Dun-qiang ZHAO Yan-xia |
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| Affiliation: | Department of Respiratory Diseases, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. lizhenxing_2006@163.com |
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| Abstract: | OBJECTIVE: To investigate the role of Rho-kinase signaling pathway in human airway smooth muscle cell (ASMCs) migration and cytoskeletal reorganization induced by endothelin-1 (ET-1). METHODS: Primary cultured human ASMCs obtained by tracheal explant culture method were examined for cell migration in response to ET-1 treatment using modified Boyden chambers. The changes in actin cytoskeletal reorganization were observed under confocal laser scanning microscope, and the phosphorylation of myosin-phosphatase target 1 (p-MYPT1) was examined using Western blot analysis. RESULTS: At the concentration of 0.1, 1, 10, and 100 nmol/L, ET-1 induced migration of the ASMCs, and 10 nmol/L ET-1 produced the most obvious effect (P<0.01). Rho-kinase inhibitor Y-27632 showed a dose-dependent inhibitory effect on ET-1-induced ASMC migration, and in cells exposed to 10 nmol/L ET-1, Y-27632 at 10 micromol/L significantly blocked ASMC migration (P<0.01). ET-1 (10 nmol/L) exposure resulted in reorganization of actin cytoskeleton and formation of stress fibers in the ASMCs, which were obviously inhibited by Y-27632. Compared with the control group, the AMSCs showed significant enhancement of p-MYPT1 protein expression after ET-1 exposure for 15 and 30 min (P<0.01), but prolonged exposure failed to result in the expression enhancement (P>0.05). CONCLUSION: Rho-kinase signaling pathway may play an important role in ET-1-induced ASMC migration and reorganization of actin cytoskeleton. |
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| Keywords: | human airway smooth muscle cells endothelin-1 migration actin cytoskeleton Rho kinase |
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