ATP缺失时大鼠近端肾小管上皮细胞骨架重组与ERM蛋白的重分布相关  

The cytoskeleton reorganization of NRK52E cells was associated with the redistribution of ERM during ATP depletion

作　　者：张利佳[1] 李卫星[2] 戈应滨[1] 宋鸣子[1] 杜军[1] 顾洛[1] 

机构地区：[1]南京医科大学生理学系,江苏南京210029 [2]泰州职业技术学院医学系,江苏泰州225300

出　　处：《南京医科大学学报（自然科学版）》2008年第1期5-8,共4页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省卫生职业技术教育课题资助(J200501)

摘　　要：目的:探讨ATP缺失时大鼠近端肾小管上皮细胞(NRK52E细胞)肌动蛋白细胞骨架重组情况及其可能机制。方法:用含0.1 μmol/L antimycin A的ATP缺失缓冲液处理培养的NRK52E细胞,建立细胞的体外ATP缺失模型;采用FITC标记的鬼笔环肽标记纤维型肌动蛋白(F-actin),用流式细胞仪检测技术分析肌动蛋白细胞骨架的重组情况;用Westernblot及免疫印迹技术检测ATP缺失后NRK52E细胞内骨架组分(Triton不溶组分)及上清组分(Triton可溶组分)中ERM(Ezrin/Radixin/Moesin)蛋白的分布改变。结果:体外ATP缺失模型建立,各实验组细胞内ATP浓度呈时间依赖性的下降(P<0.05);ATP缺失后,NRK52E细胞内纤维型肌动蛋白的量渐增,且肌动蛋白的聚合程度随ATP缺失时间延长而增加(P<0.05);ATP缺失后,ERM蛋白从细胞骨架解离进入胞浆,且ERM蛋白从细胞骨架的解离程度随ATP缺失的程度的增加而增加(P<0.05)。结论:ATP缺失后NRK52E细胞骨架重组可能与ERM蛋白的重分布相关。Objective：To investigate the cytoskeleton reorganization of NRK52E during cellular ATP depletion and its mechanisms. Methods：We established a cellular ATP depletion model was established in vitro with 0.1 μmol/L antimycin A diluted in depletion buffer;F-actin was examined with fluorescence of FITC-phalloidin by flow cytometric analyses;the expression of ERM proteins was detected through Western Blotting and immunoblotting technologies. Results：The cellular ATP depletion model was established successfully,and the cellular ATP concentration declined in a time-dependent manner（P 〈 0.05）; The F-actin of NRK52E was increasing during cellular ATP depletion, and these changes were paralleled with the severity of cellular ATP depletion （P 〈 0.05）. When ATP was depleted,the ERM protein detached from the cytoskeleton into the cytoplasma. Conclusion：The cytoskeleton reorganization of NRK52E cells might be associated with the redistribution of ERM during ATP depletion.

关 键 词：ATP缺失 肾小管上皮细胞(NRK52E) 肌动蛋白 细胞骨架 ERM蛋白 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]