二氧化硅致肺成纤维细胞连接蛋白Cx43定位的改变  

Localization of the connexin 43 gap-junction protein in silicon dioxide exposed to pulmonary fibroblasts

作　　者：毛国根[1] 曾群力[1] 叶少菁[1] 

机构地区：[1]浙江大学医学院劳动卫生与环境卫生研究所,杭州310031

出　　处：《中华劳动卫生职业病杂志》2001年第5期354-356,F004,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目 (3 9670 62 8);浙江省医药卫生优秀青年科技人才专项科研基金资助项目 (2 0 0 0QN0 0 4)

摘　　要：目的　研究SiO2 刺激肺泡巨噬细胞 (PAM)培养基上清液对肺成纤维细胞连接蛋白Cx43定位的影响 ,以深入探索SiO2 抑制肺成纤维细胞间隙连接通讯 (GJIC)的作用水平。方法　用0～ 5 0 0 μg/mlSiO2 刺激PAM和佛波酯 (TPA)诱导分化的THP 1(TPA primedTHP 1,pTHP 1)细胞 (具有PAM特性的人血单核细胞株 )的培养上清液对培养的中国仓鼠肺成纤维细胞 (CHL)作用 2 4h ,采用间接免疫荧光细胞化学法和激光共聚焦扫描显微镜 (LCSM)进行连接蛋白Cx43定位的测定。结果　正常对照组相邻细胞连接处有明亮的斑片状标记 ,聚集分布连接成线 ;SiO2 刺激PAM和pTHP 1细胞培养上清液作用的CHL细胞连接处标记斑点逐渐减少 ,Cx43蛋白标记斑点出现在胞浆内 ,呈无特异性定位状态 ;随着SiO2 剂量的增加 ,细胞内Cx43蛋白标记斑点向细胞核聚集。结论　SiO2 刺激PAM和pTHP 1细胞培养上清液可以改变肺成纤维细胞Cx43蛋白的定位。Objective To determine at what level and to what degree silicon dioxide(SiO 2)may disrupt gap junctional intercelullar communication(GJIC) mediated by connexin 43(Cx43) in pulmonary fibroblasts. Methods The supernatants from the primary cultured pulmonary alveolar macrophage(PAM) and the phorbol 12 myristate 13 acetate(TPA) primed THP 1(TPA primed THP 1,pTHP 1),a monocyte like cell line with the properties of PAM,were prepared,and then stimulated pulmonary fibroblasts(CHL) for 24 hours.The localizations of Cx43 in CHL were analyzed by indirect immunofluorescence histochemistry and laser confocal scanning microscopy (LCSM). Results The normal CHL cells displayed bright punctuate labeling and formed dashes,at regions of intercellular contact.Being exposed to supernatants from SiO 2 stimulated PAM and pTHP 1,the CHL cells retained a relative low degree of Cx43 labeling at the cell periphery,localized in cytoplasm,and the individual spot,rather than plaques,were smaller compared to normal cells.As the increase of the concentrations of SiO 2,the cells showed a different staining pattern,with clear cluster labeling within the nuclear volume. Conclusions The altered localization of the gap junctional protein Cx43 mediated by SiO 2 indicated that internalization of Cx43 may contribute to the disruption of GJIC in silicotic fibroblasts.

关 键 词：连接蛋白CX43 激光共聚焦显微镜 间接免疫荧光组织化学法 肺成纤维细胞 职业卫生 二氧化硅 

分 类 号：R363[医药卫生—病理学]