微丝聚合剂重构大鼠脊髓星形胶质细胞骨架对水通道蛋白4与K+通道蛋白4.1基因表达的影响  

Effects of gene expression of aquaporins-4 and K^+ ion channel protein-4.1 after cytoskeleton reconstruction by jasplakinolide in rat spinal cord astrocytes

作　　者：杜文佳[1] 党跃修[1] 汪玉良[1] 雷栓虎[1] 黄良增 汪静 马靖琳 安丽萍 

机构地区：[1]兰州大学第二医院骨科 [2]甘肃省骨关节病重点实验室,甘肃省兰州市730030

出　　处：《中国脊柱脊髓杂志》2014年第6期555-560,共6页Chinese Journal of Spine and Spinal Cord

基　　金：甘肃省技术研究与开发专项计划项目(编号1004TCYA028);兰州大学第二医院2010年度院内科研项目(编号YJ2010-48)

摘　　要：目的:探讨不同浓度微丝聚合剂(Jasplakinolide,JSK)对大鼠脊髓星形胶质细胞水通道蛋白4(aquaporins-4,AQP4)和K+通道蛋白4.1(potassium ion channel protein-4.1,Kir4.1)基因表达的影响。方法:分离培养原代大鼠脊髓星形胶质细胞并鉴定,采用浓度为0.05μg/ml、0.10μg/ml、0.20μg/ml、0.40μg/ml的JSK干预细胞2h。噻唑蓝比色法(3-4,5-Dimethylthiazol-2-yl-25-diphenyltetrazolium brom,MTT)检测细胞的增殖情况,激光共聚焦显微镜观察细胞骨架的变化和Real-time PCR法检测AQP4和Kir4.1 mRNA表达的变化。结果:0.05μg/ml、0.10μg/ml、0.20μg/ml和0.40μg/ml浓度的JSK在干预细胞2h时与空白对照组相比对细胞的增殖无明显差异(P>0.05),各浓度间相比无统计学意义。激光共聚焦显微镜观察0.05μg/ml、0.10μg/ml和0.20μg/ml浓度的JSK可以使得微丝空间结构发生重构,而0.40μg/ml浓度则使细胞微丝发生破坏。Real-time PCR检测显示,0.05μg/ml、0.10μg/ml、0.20μg/ml和0.40μg/ml浓度的JSK均能显著降低AQP4和Kir4.1基因的表达,并与对照组相比有统计学意义(P<0.05),且0.05μg/ml、0.10μg/ml、0.20μg/ml和0.40μg/ml浓度间相比也有统计学意义。结论:0.05μg/ml、0.10μg/ml和0.20μg/ml浓度的JSK可使大鼠脊髓星形胶质细胞微丝发生重构且可以下调AQP4和Kir4.1基因的表达。Objectives： To investigate the mRNA expression of aquaporins-4（AQP4） and potassium ion channel protein-4.1（Kir4.1） by different concentration of jasplakinolide（JSK）, a microfilament polymerization agent, in spinal cord astrocytes of rats. Methods： The primary spinal cord astrocytes from new born rats were extracted and cultured. 3-4, 5-Dimethylthiazol-2-yl-25-diphenyltetrazolium brom（MTT） was used to measure the cytotoxicity of JSK by different concentrations（0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml）. The changes of cytoskeleton were monitored under the Laser Scanning Confocal Microscopy（LSCM）. The mRNA levels of AQP4 and Kir4.1 were measured after JSK with different concentrations by real-time PCR. Results： MTT assay showed no significant difference in 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml groups compared with control group at 2h. LSCM showed that JSK of 0.05μg/m, 0.10μg/ml and 0.20μg/ml could remodel the spinal cord astrocyte cytoskeleton, nevertheless, 0.40μg/ml could damage cytoskeleton. Real-time PCR analysis showed that JSK of 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml concentrations could significantly decrease the gene expression of AQP4 and Kir4.1, which was statistically significant compared with control group（P〈0.05）. Furthermore, there were significant differences among 0.05μg/ml, 0.10μg/ml, 0.20μg/ml and 0.40μg/ml groups. Conclusion： JSK of 0.05μg/ml, 0.10μg/ml and 0.20μg/ml can remodel the cytoskeleton and decrease the mRNA expression of AQP4 and Kir4.1.

关 键 词：星形胶质细胞 微丝 增殖 

分 类 号：R683.2[医药卫生—骨科学]