CD2相关蛋白在肾脏细胞中的分布及其在足细胞中的作用  

THE DISTRIBUTION OF CD2-ASSOCIATED PROTEIN IN RENAL CELLS AND ITS POSSIBLE ROLE IN PODOCYTES

作　　者：姜华军[1] 张春[1] 朱忠华[1] 刘建社[1] 邓安国[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院肾内科,武汉430022

出　　处：《解剖学报》2008年第5期688-692,共5页Acta Anatomica Sinica

基　　金：国家自然科学基金资助项目(30500245);湖北省卫生厅科研基金资助项目(NX200510)

摘　　要：目的观察CD2相关蛋白(CD2AP)在肾脏不同细胞系中的表达分布,及其与足细胞裂孔隔膜分子nephrin和细胞骨架蛋白F-actin之间的联系。方法以DMEM培养基培养人肾小球系膜细胞(HMC)和人肾小管上皮细胞系(HK-2),RPMI 1640培养基培养条件永生化小鼠足细胞系。以RT-PCR及Western blotting方法检测足细胞内CD2AP和nephrin的表达。间接免疫荧光结合激光共焦方法观察CD2AP在HMC、HK-2、未分化及已分化足细胞中的表达情况,及CD2AP与nephrin在足细胞中的共存。直接免疫荧光结合激光共焦方法观察F-actin在足细胞的表达及其与CD2AP的共存。结果CD2AP均匀分布于HK-2及未分化足细胞的核周及胞浆,而不表达于HMC细胞。在足细胞分化过程中,CD2AP的分布发生了变化,出现向周边聚集的现象。CD2AP与足细胞裂孔隔膜分子nephrin及细胞骨架蛋白F-actin在足细胞中存在共定位关系。结论CD2AP表达于上皮来源的肾脏固有细胞。CD2AP在足细胞中的分布特点,提示CD2AP可能参与足细胞的分化过程,并与裂孔隔膜分子功能及细胞骨架调节有关。Objective To study the distribution of CD2-associated protein （CD2AP） in normal renal cell lines and its interaction with nephrin and F-actin in podocytes. Methods The human mesangial cells （HMC） and HK-2 were cultured in DMEM. Conditionally immortalized murine podoeyte cell line was cultured in RPMI 1640. The expressions of CD2AP and nephrin in podocytes were examined by RT-PCR and Western blotting. The distribution of CD2AP in HMC, HK-2, differentiated and undifferentiated podocytes was observed by laser scanning confocal microscopy. The coexistence of CD2AP with nephrin and F-actin in undifferentiated podocytes were also detected. Results CD2AP was distributed within the cytoplasm and perinulcear region of HK-2 and undifferentiated podocytes, but was absent in HMC cells. Its distribution profile changed and presented as peripheral accumulation when podocytes were put into differentiation-permissive conditions. CD2AP was located together with nephrin and F-actin in podocytes. Conclusion CD2AP can be detected in epithelial-originated renal cells. The alteration of distribution profile of CD2AP indicates it may participate in the process of podocytes differentiation and be involved in the regulation of slit diaphragm and cytoskeleton.

关 键 词：CD2相关蛋白 足细胞 肾小管上皮细胞 系膜细胞 免疫荧光 激光共焦显微镜 

分 类 号：R334.1[医药卫生—人体生理学]