RNAi技术沉默星形胶质细胞Adam10基因模型的建立  

siRNA Interfering of Astrocytic Adam10 Gene Slience

作　　者：徐姣[1] 刘扬[2] 潘邓记[1] 杨园[1] 张函[1] 熊颖[3] 张强[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院神经内科,武汉430030 [2]华中科技大学同济医学院附属同济医院神经外科,武汉430030 [3]华中科技大学同济医学院附属同济医院放射科,武汉430030

出　　处：《神经损伤与功能重建》2016年第5期377-380,384,共5页Neural Injury and Functional Reconstruction

基　　金：国家自然科学基金(No.81471230);国家自然科学基金(No.81571206)

摘　　要：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si RNA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si RNA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。Objective： To establish an in vitro model of Adam10 gene silence in astrocytes by RNAi. Methods： The purified astrocytes were isolated from neonatal SD rats cerebral cortex in fresh culture medium. The quality and purity of astrocytes were identified by glial fibrillary acidic protein （GFAP） staining. The Adam10 gene siRNA sequence were transfccted to astrocytes via Lipofectamine2000. The silence efficiency was counted via examination of the relative expression RNA and protein level of Adam10 by real time PCR and western blot respectively. Results： The Adam10 siRNA sequence was transfected into astrocytes successfully. The level of Adam10 RNA and protein in RNAi group is significantly lower than that in control groups （P〈0.05）. Conclusion： Adam10 gene silencing model can be established by RNAi technology in vitro.

关 键 词：星形胶质细胞 Adam10基因 RNA干扰 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]