【作者】 朱素敏；

【导师】 李剑波；

【作者基本信息】 重庆医科大学 ， 法医学， 2010， 硕士

【摘要】 目的建立家兔神经源性休克死亡模型,运用比较蛋白质组学筛选神经源性休克死亡组与正常对照组表达差异的蛋白质,发现潜在的神经源性休克生物标记物,为诊断神经源性休克死亡提供客观依据,探索神经源性休克死亡的机制。方法1.采用外力打击家兔颈动脉窦建立神经源性休克死亡模型。2.采用双向凝胶电泳技术分离神经源性休克死亡组及正常对照组左心室全蛋白,用PDQUEST8.0软件分析双向凝胶电泳图谱,筛选出表达显著差异的蛋白点,胰酶酶解后,用HPLC-Chip/MS进行在线分析、鉴定出差异表达蛋白质。3.利用Mascot软件在UniProtKB/SWISS-PORT数据库中筛选差异表达蛋白质,进行生物信息学分析,获取差异蛋白质相关肽段的MS/MS质谱图。结果1.成功验证了前期研究中神经源性休克死亡的动物模型。2.筛选出8个神经源性休克死亡组表达上调的蛋白质,分别是:角蛋白10、角蛋白15、α-2B肾上腺素能受体、线粒体过氧化物歧化酶、过氧化物酶增殖物激活受体δ、封闭蛋白1、锌指蛋白569、热休克蛋白70,3个表达下调的蛋白质,分别是:甲基转移酶4、膜联蛋白XIIIb、组织因子途径抑制物。结论家兔神经源性休克死亡发生了蛋白表达谱的改变,鉴定出的11种蛋白是潜在的神经源性休克生物标记物,可成为神经源性休克死亡诊断的客观依据。更多还原

【Abstract】 ObjectivesTo establish neurogenic shock death model of animal. Analysis of proteins expressed differentially between neurogenic shock death group and normal control group by comparative proteomics analysis,and provide new experimental evidences to further elucidate the death mechanism of neurogenic shock from proteomic level. In order to find latent biomarkers of neurogenic shock and to offer essential diagnosticate datas of neurogenic shock death.Methods1.To establish neurogenic shock death model by hitting on the carotid sinus in rabbits.2. We set apart the proteins in left ventricular in neurogenic shock death group and normal control group by 2-DE.The 2-DE images were analyzed with PDQuest8.0 software. Trypsin digested differentia protein spots selected. HPLC-Chip/MS was adopted to on-line analyze and indentify proteins expressed differentially. 3. Mascot software was used to search for the proteins expressed differentially in UniProtKB/SWISS-PORT, bioinformatics analysis can gain MS/MS spectrum of peptide from differentially expressed proteins.Results1.Successfully verified neurogenic shock death model of animal in our previous study.2. 8 proteins upregulated significantly in established neurogenic shock death group were identified, including keratin 10、alpha-2B adrenergic receptor、superoxide dismutase, mitochondrial precursor、peroxisome proliferator activated receptor delta、zinc finger protein -569、hsc70-interacting protein、keratin 15、claudin 1,and as well as 3 proteins downregulated significantly including methyltransferase-like protein4、annexinXIIIb、tissue factor pathway inhibitor precursor.ConclusionsThe proteins expression have changed in neurogenic shock death group .The proteins which were identified are latent biomarkers and can offer possible diagnosticate datas of neurogenic shock death.更多还原