PrP106-126诱导的N2a细胞神经毒性中神经营养因子受体p75^NTR的表达变化研究  

Altered Expression of Neurotrophin Receptor p75^(NTR) in PrP106-126-induced Neurotoxicity of N2a cells

作　　者：白瑜[1] 李玉荣[1,2] 周向梅[1] 尹晓敏[1] 赵德明[1] 

机构地区：[1]中国农业大学动物医学院,北京100193 [2]河北农业大学动物科技学院,保定071001

出　　处：《畜牧兽医学报》2009年第3期416-420,共5页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家科技支撑计划项目(2006BAD06A14);科技部项目(2005KDA21205-7)

摘　　要：神经元死亡是朊病毒病的主要病理学特征。朊蛋白多肽PrP106-126能够对神经细胞表现神经毒性,引起细胞凋亡。细胞表面蛋白神经营养因子受体p75NTR的胞外区域可以与PrP106-126结合并产生促凋亡作用。作者以小鼠成神经瘤细胞N2a为细胞模型,应用荧光定量RT-PCR和Western Blot技术,以及DNA Ladder和AnnexinV-FITC/PI双重染色流式细胞凋亡检测技术对p75NTR介导的PrP106-126神经毒性分子机制进行了研究。结果发现在PrP106-126诱导的N2a细胞凋亡过程中,p75NTR的mRNA转录水平和蛋白表达水平均显著升高,以p75NTR多克隆抗体sc-6189阻断PrP 106-126与p75NTR的相互作用后,减弱了PrP106-126诱导的N2a细胞凋亡效果。该研究揭示了PrP106-126诱导的N2a细胞毒性中p75NTR受体的表达变化,为解释朊病毒病的发病机制提供了重要数据。Neuronal death is a pathological hallmark of prion diseases. Synthetic prion peptide PrP106-126 can cause neurotoxicity and induce apoptosis in neuronal cells. Extracellular region of neurotrophin receptor p75^NTR, a cell surface protein, could bind to PrP106-126 and has apoptotic effect on neurons. Using quantitative RT-PCR, Western Blot technique, DNA Ladder assay, and flow cytometry （FCM） assay with Annexin V-FITC/PI double stained, we investigated the p75^NTR-mediated neurotoxicity by PrP106-126 in mouse neuroblastoma cells N2a. The results showed that in PrP106-126-induced apoptosis of N2a cells, p75^NTRexpression was significantly upregulated at mRNA and protein levels, and blocking the interaction of p75^NTR with PrP 106-126 by p75^NTR polyclone antibody sc-6189 attenuated the apoptotic effect induced by PrP106-126. This study showed that mRNA and protein expression of receptor p75^NTR were altered in PrP106-126- induced neurotoxicity of N2a cells, and this may partially account for the pathogenesis of prion disease.

关 键 词：朊病毒 PrP106-126 P75NTR 神经毒性 小鼠成神经瘤细胞N2a 

分 类 号：S852.659.7[农业科学—基础兽医学]