RNAi技术沉默星形胶质细胞Adam10基因模型的建立

Neural Injury and Functional Reconstruction(2016)

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Abstract
目的:建立离体星形胶质细胞Adam10基因沉默模型.方法:体外培养SD大鼠乳鼠星形胶质细胞,采用免疫荧光方法标记胶质纤维酸性蛋白(GFAP)以鉴定星形胶质细胞及其纯度.通过RNA干扰(RNAi)技术,使用转染试剂Lipofectamine2000将靶向作用于大鼠Adam10基因的siRNA序列转染至星形胶质细胞内,通过Real time-PCR和Western blotting技术在RNA水平和蛋白水平检测沉默效率.结果:靶向作用于星形胶质细胞Adam10基因的siRNA序列成功转染入星形胶质细胞,并且转染Adam10基因靶向干扰序列的星形胶质细胞Adam10基因的表达在RNA水平和蛋白水平较阴性对照组明显降低(P<0.05).结论:可以通过RNAi技术建立离体星形胶质细胞Adam10基因沉默模型.
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Key words
astrocyte,Adam10 gene,RNA interfering
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