蛋白质组学-代谢组学整合策略用于血管性抑郁小鼠模型病理机制探索的研究

发布时间：2018-04-25 13:45

本文选题：血管性抑郁 + 蛋白质组学　；参考：《第二军医大学》2017年硕士论文

【摘要】：目前,抑郁症已成为全球最主要的精神卫生问题,给社会带来了沉重的负担。血管性抑郁是抑郁症的一种典型的亚型,主要由血管性疾病或脑血管危险因素引起,患者的表现除了情绪异常低落、失眠多梦、食欲不振等典型的抑郁样消极症状外,还表现为行动功能障碍、精神运动性阻碍、思维缓慢,领悟能力也会明显减弱,严重的还会导致自杀。近年来血管性抑郁在重症抑郁症患者中所占比重显著升高,不仅严重影响患者的生活质量,对患者的工作、家庭及社会都影响重大。但其现状并不乐观,由于其功能障碍严重、成因复杂,临床诊断指标模糊,目前对于血管性抑郁的病理机制尚未有全面的阐释,基础性研究较为薄弱。因此血管性抑郁的治疗效果也不理想、疗程长且容易复发。为了积累对血管性抑郁的基础性探索,为其机制的研究、临床诊断标志物的发现以及治疗药物的开发做出一定的贡献,本研究将基于液相-质谱平台的蛋白质组学技术和代谢组学技术相结合,试图建立针对血管性抑郁小鼠模型的蛋白代谢整体调控网络,较为全面的阐释血管性抑郁对生物体的影响和扰动。为了达到上述目的,本研究采用双侧颈总动脉结扎的全脑缺血(GCI)方法来构建血管性抑郁小鼠动物模型,通过对血管性抑郁小鼠海马组织的代谢组学分析和iTRAQ标记的蛋白质组学分析,我们共筛选得到了44个差异代谢物和304个差异蛋白质。借助Ingenuity Pathway Analysis软件我们对差异代谢物和蛋白质进行了关联及整合分析,构建与血管性抑郁疾病相关联的蛋白-代谢整体调控网络。而后,我们使用LC-MS/MS,western blot和Rt-Q-PCR等技术,对生物信息学分析聚焦通路所涉及的生物分子进行了定量验证。验证结果提示机体发生了神经可塑性调节的改变、兴奋性能神经递质运输及作用的变化、神经细胞增殖与凋亡的失衡,以及氨基酸、脂质及能量代谢的紊乱。在我们的组学分析中筛选到了这些受到扰动的通路中的蛋白质及其下游代谢物。不同层次的生物分子间的关联,将为深入研究血管性抑郁的发生机制提供更有针对性以及可信度更高的着力方向。同时这些发生变化的生物分子也将成为血管性抑郁的潜在治疗靶点或生物标志物。因此,本文呈现的这种将蛋白质组学与代谢组学信息整合来准确快速地聚焦研究方向,筛选潜在靶点及生物标志物的方法和策略,对于像血管性抑郁这一类基础研究尚未展开、积累较为薄弱的疾病对象来说,可以为更深层次的研究缩小范围,提供更为精准,更为可信,更具开拓潜力的研究方向。同时,也有助于血管性抑郁疾病进一步的药物发现和临床诊断及治疗手段的开发,具有一定优势。
[Abstract]:At present, depression has become the most important mental health problem in the world, which brings a heavy burden to the society. Vascular depression is a typical subtype of depression, mainly caused by vascular diseases or cerebrovascular risk factors. It is also shown as movement dysfunction, psychomotor obstacle, slow thinking, obvious weakening of comprehension ability, and serious suicide. In recent years, the proportion of vascular depression in patients with severe depression has increased significantly, which not only seriously affects the quality of life of patients, but also has a significant impact on the work, family and society of patients. But its present situation is not optimistic, because its function obstacle is serious, the cause of formation is complex, the clinical diagnosis index is fuzzy, the pathological mechanism of vascular depression has not been fully explained at present, the basic research is relatively weak. Therefore, the treatment of vascular depression is not ideal, the course of treatment is long and easy to relapse. In order to accumulate the basic exploration of vascular depression, and to contribute to the study of its mechanism, the discovery of clinical diagnostic markers and the development of therapeutic drugs, In this study, proteomics and metabolic techniques based on liquid-mass spectrometry platform were combined to establish a global regulatory network of protein metabolism for vascular depression mice. A more comprehensive explanation of vascular depression on the biological impact and disturbance. In order to achieve the above purpose, the animal model of vascular depression in mice was established by global cerebral ischemia (GCI) with bilateral common carotid artery ligation. Through the metabolomics analysis of hippocampus and the proteomic analysis of iTRAQ labeling in the hippocampus of vascular depression mice, 44 differential metabolites and 304 differential proteins were screened out. With the help of Ingenuity Pathway Analysis software, the differential metabolites and proteins were analyzed and integrated to construct the protein-metabolism regulatory network associated with vascular depression. Then we used LC-MS / MS western blot and Rt-Q-PCR to quantitatively verify the biomolecules involved in the bioinformatics analysis of the focusing pathway. The results suggested that the changes of neuroplasticity regulation, the changes of neurotransmitter transport and action, the imbalance of nerve cell proliferation and apoptosis, and the disturbance of amino acid, lipid and energy metabolism. The proteins in these disturbed pathways and their downstream metabolites were screened in our cluster analysis. The relationship between biomolecules at different levels will provide a more specific and reliable direction for further study on the pathogenesis of vascular depression. These altered biomolecules will also be potential therapeutic targets or biomarkers for vascular depression. Therefore, the methods and strategies presented in this paper, which integrate proteomics and metabolomics information to accurately and quickly focus on the research direction, screen potential targets and biomarkers, have not been carried out for basic studies such as vascular depression. The accumulation of weak disease objects can narrow down the scope of deeper research and provide a more accurate, more reliable, more potential research direction. At the same time, it is helpful for the further discovery of drugs and the development of clinical diagnosis and treatment for vascular depression.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R749.4;R-332

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