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目的 研究SiO2 刺激肺泡巨噬细胞 (PAM)上清液对肺泡上皮细胞连接蛋白 43(Cx43)定位的影响 ,以深入探索SiO2 抑制肺泡上皮细胞间隙连接通讯 (GJIC)的作用水平。方法 不同剂量的SiO2 刺激大鼠 (SD)PAM培养上清液作用于正常貂肺泡上皮细胞株CCL 64细胞 [在体积分数为 2 %的小牛血清培养基 (RPMI1 640 )中加体积分数为 5 %的不同剂量的SiO2 刺激PAM的上清液 ]。采用间接免疫荧光的CCL 64细胞化学法和激光共聚焦扫描显微镜 (LCSM ,LeicaTCSSP)进行Cx43定位的测定。结果 正常培养的CCL 64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培养上清液作用的CCL 64细胞连接处标记斑点逐渐减少 ,Cx43标记斑点出现在胞浆内 ,呈无特异性定位状态 ,但随着SiO2 剂量的增加 ,细胞内大多数的Cx43标记斑点向细胞核聚集。结论 SiO2 刺激PAM培养上清液可以改变肺泡上皮细胞Cx43的定位。推测SiO2 抑制肺泡上皮细胞GJIC功能可能与Cx43的内移有关。
Objective To investigate the effect of SiO2-stimulated alveolar macrophage (PAM) supernatant on the location of connexin 43 (Cx43) in alveolar epithelial cells in order to explore the role of SiO2 in alveolar epithelial cell gap junctional communication (GJIC). Methods Different dosages of SiO2-stimulated rat (SD) PAM culture supernatant were applied to normal mink alveolar epithelial cell line CCL 64 [in a volume fraction of 2% fetal bovine serum (RPMI1 640) volume fraction of 5 % Of different doses of SiO2 stimulate PAM supernatant]. Cx43 localization was determined using CCL 64 cytochemistry with indirect immunofluorescence and confocal laser scanning microscopy (LCSM, Leica TCSSP). Results CCL 64 cells in normal culture had bright patch-like markers at the junctions of the cells, aggregated and distributed, and the lines were connected to each other. The CCL 64 cells with SiO2 stimulated PAM culture supernatant showed a diminished spot and the Cx43 labeled spots appeared in Within the cytoplasm, there was no specific localization, but with the increase of the dose of SiO2, most of the Cx43 labeled spots in the cells were aggregated to the nucleus. Conclusion SiO2 stimulated PAM culture supernatant can change the location of Cx43 in alveolar epithelial cells. It is speculated that the inhibitory effect of SiO2 on GJIC function of alveolar epithelial cells may be related to the internal migration of Cx43.