纤维连接蛋白HepⅡ功能域作用于体外培养的人眼小梁细胞的研究  

Experimental study on effects of heparin II domain of fibronectin on cultured human trabecular meshwork cells

作　　者：张青蔚[1] 王宁利[1] 刘旭阳[2] 陈风华[1] 王春芳[3] 吉昂[4] 刘小超[5] 

机构地区：[1]首都医科大学附属北京同仁医院北京同仁眼科中心北京市眼科学与视觉科学重点实验室,100730 [2]四川大学华西医院眼科 [3]山西医科大学第一医院眼科 [4]北京市海淀医院眼科 [5]首都医科大学附属北京同仁医院中心实验室

出　　处：《中华眼科杂志》2009年第12期1105-1110,共6页Chinese Journal of Ophthalmology

基　　金：基金项目：国家自然科学基金资助项目（30471863）

摘　　要：目的探讨纤维连接蛋白HepⅡ功能域对体外培养的人眼小梁细胞的作用。方法对照实验研究。在体外培养的人眼小梁细胞中加入纤维连接蛋白HepⅡ功能域片段，孵育18、24h，通过光镜与电镜观察小梁细胞的形态学变化，同时应用免疫荧光染色观察纤维连接蛋白HepⅡ功能域对体外培养的人眼小梁细胞肌动蛋白、纽蛋白、β-连接蛋白的影响。结果加入纤维连接蛋白HepII功能域片段10～24h，光镜下观察，可见小梁细胞逐渐发生形态学变化，表现为细胞收缩、变圆、彼此间分离；激光共聚焦显微镜下观察，呈红色细丝状着染的小梁细胞肌动蛋白染色减少；位于肌动蛋白两端绿色点、簇状着染的Vinculin染色明显减少；B—Catenin免疫荧光染色呈绿色点状，在沿细胞相连的胞膜处分布减少。电镜下观察，可见小梁细胞突起间缝隙增宽。加入纤维连接蛋白HepⅡ功能域片段24h，小梁细胞恢复至加入HepⅡ功能域片段前的状态。结论纤维连接蛋白HepⅡ功能域能够可逆性的阻断人眼小梁细胞肌动蛋白及其细胞连接。通过信号转导并重组细胞骨架肌动蛋白及其细胞连接有可能成为重要的研究方向。Objective To determine the effects of Heparin Ⅱ （ Hep Ⅱ ） domain on cultured human trabecular meshwork （HTM） ceils. Methods HTM cells were cultured and treated with Hep Ⅱ domain for 18 and 24 h. The morphological changes in HTM cells were assessed by light and electron microscopy. Changes in cell morphology and the organization of actin cytoskeleton, Vinculin, β-Catenin were assessed by using immunofluorescence. Results Treatment of Hep Ⅱ domains resulted in morphological changes from 10 to 24 h. In light microscopy, cells rounded up, retracted and detached from each other. In high performance liquid chromatography, Hep Ⅱ domains-treated cells showed that actin fibre bundles were highly concentrated at the periphery of the cells with few actiu filaments left in this area ; decreased vinculin staining was observed toward the cell periphery; decreased β-Catenin staining was also observed around the cell sub-membrane. Transmission electron microscopy showed expended intercellular space. After 24 h, changes of HTM cells were recovered. Conclusions Hep Ⅱ domains reversiblely blocks actin cytoskeleton and cell-junction of HTM cells. Disorganization of actin cytoskeleton and cell-junction in trabecular meshwork through signal transduction may be a useful strategy for the decrease of intraocular pressure.

关 键 词：纤维蛋白类 肝素 小梁网 肌动蛋白类 细胞间接合部 

分 类 号：R686[医药卫生—骨科学]