CAJ | 학술논문

目的： Nogo及Nogo-66受体（NgR）对抑制中枢神经系统再生有重要的作用。本文主要观察化学合成NgR特异性小干扰RNA（siRNA）对具有神经元分化潜能的克隆细胞系PC12细胞 NgR mRNA 和蛋白表达的影响。方法培养大鼠肾上腺嗜铬瘤细胞 PC12细胞，神经生长因子（NGF）诱导使其分化为类神经细胞，化学合成一段针对大鼠NgR基因编码区的siRNA，用阳离子脂质体使之转染类神经细胞，转染后48 h，采用实时荧光定量PCR检测mRNA水平；并采用蛋白免疫印迹检测NgR蛋白表达的变化，检测干扰的效果。结果荧光定量PCR结果显示：转染后48 h NgR mRNA水平明显下降，差异具有显著性（P＜0.05）；同时蛋白免疫印迹结果显示，NgR蛋白表达水平与对照组比较也降低，差异同样具有显著性（P＜0.05）。结论化学合成NgR特异性siRNA可以实现大鼠神经细胞内源性NgR基因沉默。
목적： Nogo급Nogo-66수체（NgR）대억제중추신경계통재생유중요적작용。본문주요관찰화학합성NgR특이성소간우RNA（siRNA）대구유신경원분화잠능적극륭세포계PC12세포 NgR mRNA 화단백표체적영향。방법배양대서신상선기락류세포 PC12세포，신경생장인자（NGF）유도사기분화위류신경세포，화학합성일단침대대서NgR기인편마구적siRNA，용양리자지질체사지전염류신경세포，전염후48 h，채용실시형광정량PCR검측mRNA수평；병채용단백면역인적검측NgR단백표체적변화，검측간우적효과。결과형광정량PCR결과현시：전염후48 h NgR mRNA수평명현하강，차이구유현저성（P＜0.05）；동시단백면역인적결과현시，NgR단백표체수평여대조조비교야강저，차이동양구유현저성（P＜0.05）。결론화학합성NgR특이성siRNA가이실현대서신경세포내원성NgR기인침묵。
Objective Nogo and Nogo-66 receptor (NgR) play important roles in inhibiting regeneration of the central nervous system. To study the expression of NgR inhibited by the chemical synthetic siRNA cognate to NgR of the cultured PC12 cells, a extensively used in vitro system for studying neurite growth. Methods PC12 cells, a rat's phaeochromocytoma cell line, was cultured and induced by nerve growth factor (NGF). A specific siRNA which was designed against NgR gene was transfected into PC12 cells with lipofectamine. Then used real-time RT-PCR and Western blot methods to detect NgR in PC12 cells. Results The results showed that the mRNA and protein level of NgR decreased. There was a significant difference compared with control set. Conclusion The specific siRNA can effectively inhibit NgR replication in cultured nervous cells.