Research on Stress Marker in the Crebrospinal Fluid and Blood by Proteomics Analysis

脑脊液和血液中应激标志物的蛋白质组学研究

基本信息

批准号：
15390221
负责人：
KAWAMURA Noriyuki
金额：
$ 8.06万
依托单位：
National Center of Neurology and Psychiatry
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15390221/
关键词：
Proteomics Stress Major Depression Psychosomatic Disorder PTSD Tsunami 津波質量分析末梢血脳脊髄液

项目摘要

Subject :We selected the research subjects according to their stress level. We used three categories when we classified the subjects. The first category is the super healthy group who have no physical diseases, no mental diseases, no medication, no history of hospitalization. The second group is the stress group but have no physical diseases, no mental diseases, no medication, no history of hospitalization. The third group is major depression patients but have no current physical diseases. We found these subjects with informed consent through screening tests and semi-structured interviews.Methods :1)Albumin and globulin were deleted from the plasma samples by blue sepharose and protein G column. The briefly purified and concentrated plasma proteins were used for direct shot gun methods and used for electrophoresis and dyed with CBB to confirm the target bands and spots. The bands and spots were also excised from the gel, digested by trypsin, and the peptides were analyzed by LCQ and identified its m/z. The data were analyzed by Mascot analysis.2)The masspectorophy data from each category of subjects were compared and stress specific peptides and depression specific peptides were selected.3)The proteins estimated by selected peptides were confirmed by western blot analysis using their specific antibodies.4)Proteinchip analysis by MALDI massspectrometry were used for stress specific patterns.Results and Discussion1)BDNF, Chromogranin A were the candidates for stress specific markers. However NK cell activities were more sensitive than these two proteins.2)Around 20 proteins were unique to the stress group.3)Hydrophobic proteinchip analysis showed stress specific patterns around 15-20KDa molecular weight areas.

主题：我们根据其压力水平选择了研究对象。当我们对主题进行分类时，我们使用了三个类别。第一类是没有身体疾病，没有精神疾病，没有药物，没有住院病史的超级健康组。第二组是压力组，但没有身体疾病，没有精神疾病，没有药物，没有住院史。第三组是主要抑郁症患者，但没有当前的身体疾病。我们通过筛查测试和半结构化访谈发现了这些受试者。方法：1）白蛋白和球蛋白被蓝色Sepharose和蛋白G柱从血浆样品中删除。短暂纯化和浓缩的血浆蛋白用于直接射击枪法，用于电泳，并用CBB染色以确认目标带和斑点。还从凝胶中切除了带和斑点，被胰蛋白酶消化，并通过LCQ分析肽并鉴定其M/Z。 2）比较了每个类别的受试者的大量数据，并选择了应力特异性肽，并选择了抑郁症特异性肽。3）3）使用其特定抗体通过蛋白质印迹分析确认了通过其特定抗体通过其特异性抗体来确认蛋白质的蛋白质Chips protine Corneinfems and pectret temert Femernf和Dembernf ins. bd ands. bd n herm yom. bd resulmssssss.应力特定标记的候选人。然而，NK细胞活性比这两种蛋白更敏感。2）大约20种蛋白质是应激组所独有的。3）疏水蛋白chip分析显示，在15-20KDA分子量面积附近的应力特异性模式。