小干扰RNA干扰大鼠神经元Nogo受体mRNA表达的实验研究  

Experiment of siRNA knocking down endogenous expression of Nogo receptor in cultured rat primary cortical and hippocampal cells

作　　者：张涛[1] 吕碧涛[2] 徐盛明[2] 徐建广[1] 袁文[2] 

机构地区：[1]交通大学附属第六人民医院骨科,上海200233 [2]第二军医大学附属长征医院骨科

出　　处：《脊柱外科杂志》2007年第4期239-241,共3页Journal of Spinal Surgery

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(small interfering RNA,siRNA)在原代神经元干扰其mRNA表达的效果。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4对化学合成的大鼠NgR特异性siRNA,72h后提取细胞总RNA,实时荧光定量RT-PCR检测NgR mRNA表达情况。结果4对siRNA均能够下调靶基因NgR的mRNA表达水平,siNgR199、siNgR562、siNgR772和siNgR964等干扰后,NgR mRNA的表达分别为对照siRNA干扰组的6.5%、62.4%、15.2%和6.86%,与对照siRNA干扰组比较有统计学意义(P<0.05)。结论化学合成的NgR特异性siRNA能够有效的干扰大鼠原代皮层和海马细胞内NgR基因mRNA的表达水平。Objective To observe the mRNA expression of Nogo receptor in rat primary cortical and hippocampal cells after the NgR-specific siRNAs chemically synthesized were transfected. Methods Four pair of NgR-specific siRNAs were selected and transfected to rat primary cortical and hippocampal cells by using TransMessenger transfection reagent. Total RNA was harvested at 72 hours posttransfection by using Trizol reagent and was tested by using real-time RT-PCR. Results The mRNA expression of Nogo receptor were suppressed by the four NgR-specific siRNAs. Compared with the control group which was transfected with scramble siRNA ,the mRNA expression of Nogo receptor treated by siNgR-199, SiNgR-562 ,SiNgR-772 and SiNgR-964 was 6.5% ,62.4% ,15.2% and 6.86% respectively, and the P values were significant compared with the control group of siRNA interfering（ P 〈0.05）. Conclusion The siRNAs specific to Nogo receptor could knock down the endogenous expression of NgR in cultured primary cortical and hippocampal neurons of rat.

关 键 词：大鼠 脊髓损伤 生长抑制物 髓磷脂蛋白质类 细胞表面受体 逆转录聚合酶链反应 

分 类 号：R651.21[医药卫生—外科学] Q78[医药卫生—临床医学]