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The roles of Weibel-Palade bodies in processing and release of endothelin-1

Weibel-Palade 小体在内皮素-1 加工和释放中的作用

基本信息

批准号：
11670029
负责人：
FUJIMOTO Sunao
金额：
$ 1.28万
依托单位：
University of Occupational and Environmental Health
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

项目摘要

1) Dynamic changes in cultured human umbilical vein endothelial cells (HUVEC) after exposure to hypoxic condition were investigated by electron microscopy, and surpernants of the cultured media ware prepared for assays of ET-1 concentration by sandwich-EIA method. A remarkable increase in number of Weibel-Palade (WP) and elevation of ET-1 concentration in the media were confirmed in the hypoxic HUVEC, when compared to the normoxic one. The rat abdominal aorta under the hypoxic condition was investigated by electron microscopy, immunocytochemistry and in-situ hybridization. The significant increase in number of WP bodies uniquely occurred in the hypoxic samples. At the same time, the degree of immunoreactivity for ET-1 and that of ET-1 mRNA signal were significantly enhanced in the hypoxic samples when compared to the normoxic ones. These findings suggest that the correlation exists between increased WP bodies and enhanced synthesis and release of ET-1 in the hypoxic endothelial cells.2 … More ) The rat thoracic aorta after cadmium administration was prepared for electron microscopy and immunocytochemistry. A remarkable increase in number of WP bodies occurred in the endothelia of cadmium-treated rat. Immunoreactivity for ET-1 and ECE-1 was preferentially localized along the endothelium control groups, and the immunoreactive intensities of ET-1 and ECE-1 were significantly enhanced in the cadmium-treated groups. By immunoelectron microscopy, immunoreactive gold particles for ET-1 were preferentially localized in WP bodies and those for ECE-1 were in WP bodies as well as along the endothelial cell membrane in control groups. By cadmium administration, immunoreactivity for ET-1 and ECE-1 appeared to be enhanced, and immunoreactivity for ET-1 in WP bodies, which are closely associated with the endothelial cell membrane became more pronounced. Thus, the present findings suggest that the correlation exists between increased WP bodies and enhanced release of ET-1, which is processed by ECE-1 in WP bodies, in a manner of a regulated release by cadmium administration. Less

1)用电镜观察了培养的人脐静脉内皮细胞（HUVEC）缺氧后的动态变化，并制备上清液，用ELISA法测定内皮素（ET-1）浓度。与常氧组相比，缺氧组HUVEC中的韦贝尔-帕拉德（WP）细胞数显著增加，培养液中ET-1浓度显著升高。应用电镜、免疫细胞化学和原位杂交技术对低氧条件下大鼠腹主动脉进行观察。WP机构的数量的显着增加，唯一发生在缺氧样品。同时，缺氧组大鼠脑组织ET-1的免疫反应性及ET-1 mRNA的表达水平均显著高于常氧组。提示缺氧时内皮细胞内WP小体增多与ET-1的合成和释放增加有关。 ...更多信息取染镉大鼠胸主动脉进行电镜和免疫细胞化学观察。镉染毒大鼠内皮细胞内WP小体数量明显增多。ET-1和ECE-1的免疫反应主要沿着内皮细胞，镉处理组ET-1和ECE-1的免疫反应强度明显增强。免疫电镜观察显示，ET-1免疫反应阳性金颗粒主要定位于WP小体，ECE-1免疫反应阳性金颗粒主要定位于WP小体，并沿内皮细胞膜沿着。通过镉的管理，ET-1和ECE-1的免疫反应性似乎增强，和ET-1的免疫反应性WP机构，这是密切相关的内皮细胞膜变得更加明显。因此，目前的研究结果表明，增加WP机构和ET-1的释放增强，这是由ECE-1在WP机构处理，在镉管理的调节释放的方式之间存在的相关性。少