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
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江苏省卫生职业技术教育课题资助(J200501)


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

    Abstract:

    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.

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张利佳,李卫星,戈应滨,宋鸣子,杜 军,顾 洛. 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[J].南京医科大学学报(自然科学版),2008,28(1):5-8

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  • 收稿日期:2007-07-19
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