利用慢病毒载体建立一种Alzheimer's症体外模型  

Lentivirus transduction of APP Swedish in primary cultured rat Hippocampal neurons

作　　者：叶旋[1] 泰文娇 孙士鹏[3] 张丹[4] Xuan Ye Wenjiao Tai Shipeng Sun Dan Zhang(Center for Drug Evaluation, China Food and Drug Administration, Beijing 100038, China Department of Molecular Biology, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA Clinical Laboratories, Guang＇ anmen Hospital, China Academy of Chinese Medical Sciences, Beijing100053, China State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences ＆ Peking Union Medical College ,Beijing 100050, China)

机构地区：[1]国家食品药品监督管理总局药品审评中心,北京100038 [2]Texas大学西南医学中心分子生物系,dallas美国75390 [3]中国中医科学院广安门医院检验科,北京100053 [4]中国医学科学院北京协和医学院药物研究所,北京100050

出　　处：《生物技术》2016年第5期495-500,共6页Biotechnology

基　　金：国家重点基础研究发展计划项目("973"计划;No.2011CB504105);教育部新世纪优秀人才项目(No.3332013128)

摘　　要：[目的]以大鼠原代培养的海马神经元为基础,建立一种阿尔兹海默症(Alzheimer's Disease,AD)体外疾病模型。[方法]利用分子克隆技术,构建带有Swedish突变的AD致病相关基因APP的慢病毒载体系统,将其导入原代培养的大鼠海马神经元。[结果]在APP野生型中了引入Swedish突变,成功构建APPSwe慢病毒载体系统,并在大鼠原代海马神经元中实现高表达。[结论]慢病毒系统能够将阿尔兹海默症致病相关基因Swedish突变APP导入大鼠原代海马神经元中,并且与对照组相比APP表达量为263.9±18.3%,为进一步研究阿尔兹海默症的致病机制以及寻找可能的治疗手段建立了一种体外评价模型。[ Objective] To build a cellular model in vitro for Alzheimer＇s Disease （AD） based on primary cultured rat hippocampal neurons. [ Methods] Lentiviral vector was used to deliver APP Swedish mutation which is considered to be one of the genetic factors for AD. APP Swedish mutation was transduced into primary cultured rat hippocampal neurons by Lentiviral delivery system. [ Results ] Point mutation for APP Swedish was generated by PCR. Lentiviral vectors carrying APP Swedish was constructed and the conditions for Lentirirus packing had been optimized. APP Swedish mutation was transduced and expressed robustly in primary cultured rat hippocampal neurons. [ Conclusion ] A cellular model for AD based on primary cultured hippocampal neurons had been built.

关 键 词：慢病毒载体 原代海马神经元 阿尔兹海默症 APP Swedish突变 

分 类 号：Q933-331[生物学—微生物学]