siRNA干扰大鼠神经元Nogo受体mRNA表达的时程研究  

siRNA knocks down mRNA expression of Nogo receptor in cultured rat neurons

作　　者：张涛[1] 袁文[1] 刘百峰[1] 曹莉[2] 肖林[2] 陈公星[3] 

机构地区：[1]第二军医大学长征医院骨科,上海市200003 [2]第二军医大学神经生物学教研室,上海市200433 [3]浙江大学医学院附属第二医院肿瘤研究所,杭州市310009

出　　处：《中国脊柱脊髓杂志》2005年第10期588-591,i0004,共5页Chinese Journal of Spine and Spinal Cord

基　　金：国家自然科学基金资助项目(30471757)

摘　　要：目的:观察大鼠Nogo受体(NgR)特异性小干扰RNA(smallinterferingRNA,siRNA)在原代神经元干扰其mRNA表达的时程效果。方法:体外培养大鼠原代皮层和海马细胞,应用阳离子脂质体转染试剂转染针对2个基因片段(199和964位点)的大鼠NgR特异性siRNA和对照siRNA,分别于转染后24h、48h、72h和96h,应用实时荧光定量PCR检测NgRmRNA表达情况。结果:2对siRNA(siNgR199和siNgR964)均能够下调靶基因mRNA的表达水平,24h、48h、72h和96h,NgRmRNA表达分别为对照siRNA组的38.12%、12.47%、3.96%、18.4%(siNgR199)和49.54%、24.25%、13.17%、37.93%(siNgR964),与对照siRNA组比较有统计学意义(P<0.05)。结论:NgR特异性siRNA能够在原代皮层和海马细胞下调靶基因mRNA的表达水平,且基因沉默效果在转染后3d最显著。Objective.To observe the mRNA expression of Nogo receptor（NgR） in rat primary cortical and hippocampal cells at different timepoint after the NgR-specific siRNAs were transfected.Metbod：Two NgR- specific siRNAs were selected and transfected into rat primary cortical and hippocampal cells by using transmessenger transfection reagcnt.Total RNA was harvested at 24h,48h,72h and 96h posttransfection by using Trizol reagent and was tested by using real-time PCR.Result：The mRNA expression of NgR was suppressed by the two NgR-specific siRNAs tested.Compared with the control group which was transfected with scramble siRNA,the mRNA expression of NgR interfered by siNgR199 and siNgR964 was 38.12%,12.47%,3.96%, 18.4%（siNgR199） and 49.54%,24.25%,13.17%,37.93%（siNgR964） at different timepoint respectively,and the P values were significant compared with the control siRNA （P〈0.05）.Conclusion：The siRNAs specific to NgR could knock down the endogenous expression of taget gene,which is most effective at 3th day posttransfection in cultured cells.

关 键 词：NOGO受体 RNA干扰 基因沉默 实时荧光定量PCR 

分 类 号：Q786[生物学—分子生物学] R683.2[医药卫生—骨科学]