CAJ | 학술논문

目的建立局灶性缺血性脑梗死模型,观察Nogo-66受体(NgR1)拮抗剂(sNgR1-Fc)促进内源性成体神经前体细胞(NPCs)定向分化的作用,并探讨其机制.方法取SD大鼠12只,光化学法建立大脑皮层局灶缺血梗死(PCI)模型.设假手术组、PBS组和sNgR1-Fc治疗组.在PCI术后第1～3天,通过小型渗透性微泵持续性地向梗死灶同侧的侧脑室灌注sNgR1-Fc或同等体积的PBS;第4～6天,通过腹腔注射BrdU(bromodeoxyuridine)；在PCI术后35 d,利用免疫组化染色观察各组海马齿状回NeuN+/BrdU+细胞的比率；Western blot检测梗死侧海马Notch1、Mash1及Neuro D蛋白的表达情况.结果光化学法可以成功地建立局灶性缺血性脑梗死模型.PCI术后35 d时梗死灶同侧的海马齿状回,sNgR1-Fc处理组的BrdU+细胞的数目显著高于PBS组(P＜0.01),NeuN+/BrdU+细胞的比率也显著高于PBS组(P＜0.05).Western blot实验表明,sNgR1-Fc治疗组海马的Notch1、Mash1、Neuro D蛋白的表达显著高于PBS组,后者又显著高于假手术组.结论 NgR1拮抗剂NgR1-Fc能够促进脑梗死后大脑中的神经前体细胞向神经元方向分化,这一作用可能是通过影响Notch和bHLH家族信号通路而发挥的.
목적 건립국조성결혈성뇌경사모형,관찰Nogo-66수체(NgR1)길항제(sNgR1-Fc)촉진내원성성체신경전체세포(NPCs)정향분화적작용,병탐토기궤제.방법 취SD대서12지,광화학법건립대뇌피층국조결혈경사(PCI)모형.설가수술조、PBS조화sNgR1-Fc치료조.재PCI술후제1～3천,통과소형삼투성미빙지속성지향경사조동측적측뇌실관주sNgR1-Fc혹동등체적적PBS;제4～6천,통과복강주사BrdU(bromodeoxyuridine)；재PCI술후35 d,이용면역조화염색관찰각조해마치상회NeuN+/BrdU+세포적비솔；Western blot검측경사측해마Notch1、Mash1급Neuro D단백적표체정황.결과 광화학법가이성공지건립국조성결혈성뇌경사모형.PCI술후35 d시경사조동측적해마치상회,sNgR1-Fc처리조적BrdU+세포적수목현저고우PBS조(P＜0.01),NeuN+/BrdU+세포적비솔야현저고우PBS조(P＜0.05).Western blot실험표명,sNgR1-Fc치료조해마적Notch1、Mash1、Neuro D단백적표체현저고우PBS조,후자우현저고우가수술조.결론 NgR1길항제NgR1-Fc능구촉진뇌경사후대뇌중적신경전체세포향신경원방향분화,저일작용가능시통과영향Notch화bHLH가족신호통로이발휘적.
Objective To study the effect of neuronal Nogo-66 receptor (NgR1) antagonist,soluble Nogo-66 receptor (sNgR1-Fc),on promoting the endogenous neural precursor cells (NPCs) differentiating into neurons in order to clarify the mechanism.Methods The cortical infarction was induced by photochemistry,named photothrombotic cortical injury (PCI).Twelve Sprague Dawley rats were randomly divided (random number) into three groups:Sham-operated group,PBS group,and sNgR1-Fc group.PBS (PBS group) or sNgR1-Fc (sNgR1-Fc group) was injected into the lateral ventricle of brain with a minipump.BrdU (Bromodeoxyuridine) was injected into the peritoneal cavity 4-6 days after PCI.The subdentate gyrus zone (SGZ) of brain from sacrificed rat was harvested for Immunohistochemistry to observe the ratio of NeuN +/BrdU + cells 35 days after PCI.Proteins including Nestin、Notch1 and Mash1 were detected by Western Blot.Results The cortical infarction in rat was successfully induced by photochemistry.Thirty-five days after PCI,the BrdU + cells number and theratio of NeuN +/BrdU + in the SGZ of the ipsilateral cerebrum hemisphere with PCI were significantly higher in sNgR1-Fc group than those in PBS group (P ＜ 0.05).The levels of Notch1,Mash1 and Neuro D in the sNgR1-Fc group were significantly higher than those in the PBS group (P ＜ 0.05),which were significantly higher than those in the Sham-operated group.Conclusions sNgR1-Fc could promote the endogenous NPCs differentiating into neurons in a cortical infarction model.The mechanisms may be attributed to the Notch/bHLH (proneural basic helix-loop-helix genes) signaling way.