《中国康复理论与实践》

《中国康复理论与实践》 ›› 2014, Vol. 20 ›› Issue (7): 616-620.

• 论文 • 上一篇    下一篇

细胞松弛素D对大鼠脊髓星形胶质细胞水通道蛋白和内向整流性钾通道4.1 基因表达的影响

杜文佳，汪玉良，党跃修，雷栓虎，黄良增，汪静，马靖琳，安丽萍   

  1. 1.兰州大学第二医院骨科，甘肃兰州市730030；2.甘肃省骨关节病重点实验室，甘肃兰州市730030。
  • 收稿日期:2013-10-22 修回日期:2013-11-27 出版日期:2014-07-25 发布日期:2014-07-25
  • 通讯作者: 汪玉良

Effects of Cytochalasin D on Expression of Aquaporins and Inward Rectifying Potassium Channel 4.1 Gene in Spinal Cord Astrocytes of Rats

DU Wen-jia, WANG Yu-liang, DANG Yue-xiu, et al.   

  1. Department of Orthopedics, The Second Hospital of Lanzhou University,Orthopaedics Key Laboratory of Gansu Province, Lanzhou, Gansu 730030, China
  • Received:2013-10-22 Revised:2013-11-27 Published:2014-07-25 Online:2014-07-25

被引次数

7

摘要： 目的探讨不同浓度细胞松弛素D(CytD)对大鼠脊髓星形胶质细胞骨架重构，水通道蛋白(AQP)1、AQP4 及内向整流性钾通道4.1(Kir4.1)基因表达的影响。方法原代培养大鼠脊髓星形胶质细胞，加CytD 0.05 μg/ml、0.10 μg/ml、0.20 μg/ml、0.40μg/ml、0.80 μg/ml 和1.00 μg/ml 共培养2 h、12 h 和24 h。MTT法检测细胞增殖情况；鬼笔环肽联合Hoechst 33342 套染后激光共聚焦显微镜下观察共培养2 h 后细胞骨架重构情况；RT-PCR法检测共培养2 h 时AQP1、AQP4 和Kir4.1 mRNA表达水平。结果细胞生存率随CytD 浓度升高和作用时间延长而减少。CytD 使大鼠脊髓星形胶质细胞微丝发生解聚、弯曲，但极性未失。CytD 0.05μg/ml、0.10 μg/ml、0.20 μg/ml 和0.40 μg/ml 上调AQP1、AQP4 和Kir4.1 mRNA表达。结论适当浓度CytD可以重建星形胶质细胞骨架，上调大鼠脊髓星形胶质细胞AQP1、AQP4和Kir4.1基因表达。

关键词: 星形胶质细胞, 细胞松弛素, 细胞骨架, 水通道蛋白, 内向整流性钾通道, 脊髓, 大鼠

Abstract: Objective To investigate the expression of aquaporin (AQP) 1, AQP4, inward rectifying potassium channel 4.1 (Kir4.1) and cytoskeleton features of rat spinal cord astrocytes after cytochalasin D (CytD) intervention. Methods Spinal cord astrocytes isolated from 2~3-day-old rats were cultured till confluency. MTT was used to assess survival rate of astrocytes 2 h, 12 h and 24 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD, respectively. Confocal microscopy was used to observe cytoskeleton features of astrocytes 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml of CytD. The expression of AQP1, AQP4, Kir4.1 mRNA were determined with real-time PCR 2 h after co-cultured with 0.05 μg/ml, 0.10 μg/ml, 0.20 μg/ml, 0.40 μg/ml, 0.80 μg/ml and 1.00 μg/ml of CytD. Results The survival rate of rat spinal cord astrocytes reduced with the time of co-culture and concentration of CytD (P<0.05). The cytoskeleton of astrocytes was reconstructed. The expression of AQP1, AQP4 and Kir4.1 mRNA increased after co- cultured with 0.05~0.40 μg/ml of CytD. Conclusion The appropriate dosage of CytD may remodel the cytoskeleton and increase the mRNA expression of AQP1, AQP4 and Kir4.1 in spinal cord astrocytes of rats.

Key words: astrocyte, cytochalasin, cytoskeleton, aquaporin, inward rectifying potassium channel, spinal cord, rats