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

发布时间：2018-06-29 04:42

  本文选题：星形胶质细胞 + 细胞松弛素　； 参考：《中国康复理论与实践》2014年07期

【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 0.05μg/ml、0.10μg/ml、0.20μg/ml、0.40μg/ml、0.80μg/ml和1.00μg/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、0.10μg/ml、0.20μg/ml和0.40μg/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。
[Abstract]:Objective to investigate the effects of different concentrations of cytochalasin D (CytD) on cytoskeleton remodeling, aquaporin (AQP) 1, AQP4 and inward rectifier potassium channel 4.1 (Kir4.1) gene expression in rat spinal astrocytes. Methods Primary cultured rat spinal astrocytes were cultured with CytD 0.05 渭 g / ml 0.10 渭 g / ml 0.20 渭 g / ml 0.40 渭 g / ml / ml 0.80 渭 g/ml and 1.00 渭 g/ml for 2 h, 12 h and 24 h, respectively. The cytoskeleton remodeling after 2 h of co-culture was observed under a set of hoechst 33342 sets of laser scanning confocal microscopy. RT-PCR was used to detect the expression levels of AQP1, AQP4 and Kir4.1 mRNA at 2 h after co-culture. Results the cell survival rate decreased with the increase of CytD concentration and the duration of action. CytD induced depolymerization and curvature of microfilaments in rat spinal astrocytes, but the polarity was not lost. CytD 0. 10 渭 g / ml, 0. 20 渭 g/ml and 0. 40 渭 g/ml upregulated the expression of AQP1AQP4 and Kir4.1 mRNA. Conclusion the appropriate concentration of CytD can reconstruct the astrocytoskeleton and up-regulate the expression of AQP1-AQP4 and Kir4.1 genes in rat spinal astrocytes.
【作者单位】： 兰州大学第二医院骨科;甘肃省骨关节病重点实验室;
【基金】：甘肃省技术研究与开发专项计划项目(No.1004TCYA028) 兰州大学第二医院2010年度院内科研项目(No.YJ2010-48)
【分类号】：R338
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本文编号：2080899