【摘要】 目的测定氯化锰（MnCl₂）处理脉络丛上皮细胞质Z310后升高的抑制素蛋白1（PHB1）在亚细胞中的分布及重定位,以及在细胞胞浆中的主要定位亚细胞器,并观察PHB1与细胞生长调控蛋白P53在细胞内是否有共定位关系。方法采用免疫蛋白印迹法（Western Blot）测定MnCl₂（0,100,200μmol/L）处理24 h后Z310细胞胞浆和胞核内的PHB1表达的变化及其重定位;采用免疫荧光法（Immunofluorescence,IF）结合激光共聚焦显微镜,检测并观察MnCl₂处理后PHB1的亚细胞空间分布,及其与P53的亚细胞共定位特征。结果 Western Blot检测表明,与对照组比较,MnCl₂处理后Z310细胞内PHB1的表达在胞浆中升高,同时在胞核内降低（P<0.05）;免疫荧光法显示MnCl₂处理后PHB1在胞浆和胞核均存在,且在胞浆中主要分布于线粒体上;在胞浆和胞核内发现PHB1与P53有共定位（Co-localization）。结论MnCl₂处理后,上调表达的PHB1蛋白主要位于胞浆,同时发现胞核内PHB1降低,提示MnCl₂能够引起Z310细胞中PHB1蛋白升高并从胞核向胞浆移动,这种亚细胞水平的PHB1空间重定位（translocation）改变是锰毒性依赖性的;胞浆和胞核中的PHB1均与P53有共定位关系;胞浆中的PHB1主要存在于线粒体上。更多还原

【Abstract】 Objective To clarify the sub-cellular distribution and localization of PHB1 up-regulated by MnCl₂. In addlition,the main localization on organelle in cytoplasm and the co-localization with cell growth regulation protein P53 were included. Methods Western Blot was used to determine the PHB1 expression in cytoplasm and nucleus in Z310 which was treated with MnCl₂（ 0,100,200 μmol/L） for24 h. And based on laser scanning confocal microscopy,the immune fluorescence（ IF） was conducted to confirm the subcellular localization of PHB1 as well as the co-localization with P53 after treating with MnCl₂. Results The expression of PHB1 was significantly elevated in cytoplasm,while decreased in nucleus by MnCl₂（ P < 0. 05）. IF staining showed PHB1 were present in nucleus and cytoplasm and mainly locates on mitochondrial in cytoplasm. Also,there exists co-localization between PHB1 and P53. Conclusion The up-regulation of PHB1 mainly exists in cytoplasm and the decrease in nucleus,which was dose-dependent. It indicates PHB1 translocates from nucleus to cytoplasm after treating with MnCl₂. The co-localization between PHB1 and P53 exits in cytoplasm and nucleus while PHB1 mainly locates on mitochondrial in cytoplasm.更多还原