脑缺血大鼠脑组织和血浆的蛋白组学分析及STVNa对脑缺血损伤的调节作用

发布时间：2018-07-08 18:58

本文选题：脑缺血 + 蛋白组学　；参考：《华南理工大学》2015年硕士论文

【摘要】：缺血性脑卒中是死亡率最高的第三大疾病。其病理变化的信号传导通路尚不明确,本文采用基于LC-MS/MS的高通量鸟枪法蛋白组学研究方法对局灶性脑缺血大鼠的血浆和脑组织中的蛋白进行测定,并对涉及不同信号通路的相关蛋白进行了鉴定和分析。此外,异甜菊醇钠(isoteviol sodium,STVNa)被证实具有脑保护作用,我们同时研究了STVNa对上述通路变化的调控作用。研究内容主要如下:将雄性成年Sprague-Dawley大鼠分为正常组、脑缺血(MCAO)组及缺血给药(STVNa)组,分别收集脑组织胞浆及血浆。建立基于阳离子交换的蛋白分级方法和基于LC-MS/MS的蛋白鉴定方法。1)分析正常组与缺血组的血浆中特异性表达的蛋白。质谱鉴定到176个只在正常组特异性表达的蛋白,和200个只在缺血组特异性表达的蛋白。进一步对缺血组中特异性表达的蛋白进行基因表达组织特异性分析,得到6个具有高度脑组织特异性的蛋白。2)分析正常组与缺血组的脑组织中特异性表达的蛋白。质谱鉴定到648个只在正常组特异性表达的蛋白,和147个只在缺血组特异性表达的蛋白。进一步对缺血组中特异性表达的蛋白进行通路分析,发现大量与炎症、细胞死亡及自我修复相关的通路。3)分析正常组、缺血组及给药组的脑组织中鉴定到的蛋白。质谱鉴定到279个只在给药组中特异性表达的蛋白。通过进一步的通路分析发现,STVNa抑制了脑缺血后炎症通路的增强和细胞生长通路的降低,同时促进碳水化合物代谢通路与脂肪酸代谢通路。结论:1)在缺血大鼠血浆中鉴定到6个具有高度脑特异性的蛋白可为脑卒中生物标志物的筛选提供基础;2)脑缺血后(4h)脑组织中与炎症、细胞凋亡、自我修复相关的通路相对活跃;3)STVNa能抑制炎症、促进细胞生长、促进代谢重构,在脑缺血损伤中发挥调节作用。
[Abstract]:Ischemic stroke is the third most fatal disease. The signal transduction pathway of its pathological changes is not clear. In this paper, the protein in plasma and brain tissue of rats with focal cerebral ischemia was determined by high-throughput birdshot proteomics based on LC-MS / MS. The related proteins involved in different signal pathways were identified and analyzed. In addition, sodium isoteviol sodiferol (STVNa) has been proved to have brain protective effect. We also studied the regulation of STVNa on the changes of these pathways. The main contents of the study were as follows: adult Sprague-Dawley rats were divided into normal group, cerebral ischemia (MCAO) group and ischemic administration group (STVNa). A cationic exchange based protein classification method and a LC-MS / MS protein identification method were established to analyze the specific expression of proteins in the plasma of normal and ischemic groups. A total of 176 proteins specifically expressed in normal group and 200 in ischemic group were identified by mass spectrometry. Furthermore, six proteins with high brain tissue specificity were obtained by tissue specific analysis of the specific expression of the proteins in the ischemic group and in the ischemic group. (2) the specific expression of the protein in the brain tissue of the normal group and the ischemic group was analyzed. 648 proteins were specifically expressed in normal group and 147 proteins in ischemic group were identified by mass spectrometry. A large number of pathways related to inflammation, cell death and self-repair were found to analyze the proteins identified in brain tissues of normal group, ischemic group and administration group. A total of 279 proteins were identified by mass spectrometry and expressed only in the administration group. It was found that STVNa inhibited the increase of inflammatory pathway and the decrease of cell growth pathway after cerebral ischemia, and promoted carbohydrate metabolism pathway and fatty acid metabolic pathway. Conclusion: 1) six highly brain-specific proteins were identified in the plasma of ischemic rats to provide a basis for the screening of biomarkers for stroke. 2) inflammation and apoptosis were observed in the brain tissue (4 h after cerebral ischemia). STVNa can inhibit inflammation, promote cell growth, promote metabolic remodeling, and play a regulatory role in cerebral ischemia injury.
【学位授予单位】：华南理工大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R743.3

【参考文献】