寡核苷酸对星形胶质细胞硫酸软骨素蛋白聚糖表达调控的实验研究

Express regulatory of ODN-PSs on astrocyte CSPGs

目的用人工合成的内切磷硫酰寡脱氧核糖核苷酸(ODN-PSs)对星形胶质细胞进行体外干预培养,分析硫酸软骨素蛋白聚糖表达,探讨ODN-PSs与神经损伤修复的关系。方法采用星形胶质细胞体外诱导培养技术,制备得到ODN-PSs条件培养的细胞;利用斑点杂交、免疫印迹和RT-PCR方法对细胞表达的硫酸软骨素蛋白聚糖和木糖基转移酶-1(XT-1)mRNA进行分析。结果制备的星形胶质细胞采用抗GFAP抗体进行免疫细胞化学染色法进行鉴定,纯度在95%以上;条件培养星形胶质细胞的免疫印迹和RT-PCR分析结果表明,XT-1 ODN-PSs可减少星形胶质细胞CSPG蛋白和XT-1基因的表达。结论人工合成XT-1 ODN-PSs寡核苷酸对星形胶质细胞硫酸软骨素蛋白聚糖表达有明显的抑制作用,从而有利于损伤神经核突起的再生。

Objective Use synthetic phosphorothioate oligonucleotid（ODN-PSs） interfere astrocyte in vitro , analysis the express of chondroitin sulfate proteoglycans （CSPGs）. Methods In vitro cell culture method was adopted, use dot-blot hybridization, western blot and RT-PCR to analysis the mRNA of CSPGs and Xylosyltransferase-1（XT- 1）. Result Use Immunocytochemical stain to identify the astrocyte, and find the purity of glial fibrillary acidic protein （CSPG） is more than 95 %. TheResults of western blot and RTPCR show that ODN-PSs can reduce the level of mRNA of CSPGs and XT-1. Conclusion ODN-PSs can significantly inhibit the express of CSPGs,so as to benefit the regeneration of nucleus.