aqp4基因多态性与视神经脊髓炎遗传易感性的相关研究

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1、贵阳医学院2012届硕士研究生论文《中国图书资料分类法》单位代码：10660分类号：R744.5学号：S090264贵阳医学院2012届临床医学硕士专业学位论文（临床论文部分）AQP4基因多态性与视神经脊髓炎遗传易感性的相关研究研究生：王青松导师：楚兰教授年级：2009级专业：神经病学2012年05月08日0贵阳医学院2012届硕士研究生论文目录摘要„„„„„„„„„„„„„„„„„„„„„„„„2缩略词表„„„„„„„„„„„„„„„„„„„„„„„3前言„„„„„„„„„„„„„„„„„„„„„„„„4材

2、料与方法„„„„„„„„„„„„„„„„„„„„„„6结果„„„„„„„„„„„„„„„„„„„„„„„„12讨论„„„„„„„„„„„„„„„„„„„„„„„„20结论„„„„„„„„„„„„„„„„„„„„„„„„24参考文献„„„„„„„„„„„„„„„„„„„„„„„25致谢„„„„„„„„„„„„„„„„„„„„„„„„28英文摘要„„„„„„„„„„„„„„„„„„„„„„„29论文原创性声明„„„„„„„„„„„„„„„„„„„„31综述„„„„„„„„„„„„„„„„„„„„„„„„32在读期

3、间发表的论文„„„„„„„„„„„„„„„„„„40附：临床能力考核有关材料：临床轮转考核表(按时间顺序)学术活动情况表(按时间顺序)临床能力考试及学位答辩情况表1贵阳医学院2012届硕士研究生论文AQP4基因多态性与视神经脊髓炎遗传易感性的相关研究专业：神经病学研究生：王青松导师：楚兰教授摘要目的：初步探讨AQP4外显子区域基因多态性与我国视神经脊髓炎（NMO）患者遗传易感性的关系，根据NMO患者单核苷酸多态性（SNP）位点的相关信息，通过我们建立的不同位点突变的NMO-IgG检测细胞株进行检测，从而对不同

4、研究组总体的NMO-IgG抗体滴度水平的差异来分析不同位点的突变可能产生的相应的蛋白质的功能变化，进而从AQP4基因水平这一角度来分析探讨NMO的发病机制。方法：对36例符合入选标准的NMO患者以及34例多发性硬化（MS）、30例脑梗死（CI）、30例正常对照组（NC）为研究对象。用常规酚/氯仿法从患者及正常对照组全血标本中提取基因组DNA，应用PCR扩增AQP4外显子区基因（包括AQP4-exon1和AQP4-exon5），再对PCR产物进行纯化及正反双向测序，再通过与GeneBank公布的人类AQP4基因

5、序列比较，寻找阳性突变位点，并比较各阳性突变位点在NMO、MS、CI、正常对照组中突变率的差异。后通过特异性的定点诱变技术对pEGFP-N1-AQP4质粒诱导相应位点的突变生成相应的突变AQP4质粒，采用脂质体转染法,将相应的质粒转染进入HEK-293T细胞稳定表达后进行NMO-IgG抗体滴度的检测。应用SPSS11.5统计软件进行相关数据分析。结果：我们发现视神经脊髓炎NMO组其5号外显子区域有单核苷酸多态性位点4个，突变的位点为C9414T、G9416C、A9458C、C9524A，而多发性硬化组，脑梗死

6、组及正常对照组均未发现相应的SNP位点，同时NMO患者1号外显子区域未发现SNP位点。不同的SNP位点突变细胞株进行相关检测后发现，组间单核苷酸多态性位点的分布的差异有统计学意义（P<0.05）,组间血清抗体滴度水平具有统计学差异（P<0.05）。结论：我们成功地完成了AQP4基因外显子区域的PCR扩增和DNA序列测定，共发现了4个AQP4-exon5阳性突变位点，推测AQP4基因编码区的变异或多态性可影响靶蛋白AQP4的表达，进而可能参与NMO的发病机制。关键词:视神经脊髓炎;单核苷酸多态性;AQP4基因；

7、多发性硬化2贵阳医学院2012届硕士研究生论文缩略词表缩略词英文全称中文全称NMONeuromyelitisOptica视神经脊髓炎MSMultipleSclerosis多发性硬化AQP4Aquaporin-4水通道蛋白4HR-NMOHigh-riskSyndromesforNMO视神经脊髓炎高危综合征HEK293THumanEmbryonicKidney293T人胚肾293细胞EDSSExpandedDisabilityStatusScale扩展残疾状况评分SNPSingleNucleotidePolymo

8、rphism单核苷酸多态性CNSCentralNervousSystem中枢神经系统CICerebralInfarction脑梗死NCNormalControl正常对照GFPGreenFluorescentProtein绿色荧光蛋白PCRPolymeraseChainReaction聚合酶链反应3贵阳医学院2012届硕士研究生论文1.前言视神经脊髓炎（neuromyelitisoptica,NMO）