低氧预适应抗缺氧神经损伤的物质基础及其调控网络的探索表征

Though it is well recognized that hypoxic preconditioning exerts endogenous neuroprotective effect against severe hypoxia, the mediators for the protective effect is still not fully understood. Based on the previous studies, this study is about to apply the proteomics and metabolomics technologies to explore the differential proteins and metabolites at different hypoxic exposure times during hypoxic preconditioning on acute repetitive hypoxic mice model. Then, protein interaction databases, database of metabolic networks and systems biology network modeling techniques are used to build the protein expression and metabolic regulatory network triggered by hypoxic preconditioning, resolve the key nodes associated with neuroprotective effect of hypoxic preconditioning, so as to obtain hypoxic preconditioning-induced potential key proteins or metabolites against hypoxic neuronal damage. Finally, by downregulating or upregulating these key regulatory proteins or metabolites by siRNA or adding inhibitor or agonist,to verify their effect in hypoxia adaptation pathway and mechanism in the regulatory network at the cellular level, so as to interpret the mediators and regulatory mechanism of hypoxic preconditioning against hypoxic neuronal damage in the molecular network level. By this study, we try to explore key regulatory molecules with the activity of anti-hypoxic neuronal damage and new drug targets with independent intellectual property rights, and to provide a theoretical basis for the development of innovative anti-hypoxia drug candidates.

低氧预适应对缺氧神经损伤的内源性保护作用已有共识，但对其产生的内源性保护作用的物质基础认识仍处于初级阶段。本项目拟在既往研究的基础上，整合蛋白质组学和代谢组学技术，以急性重复低氧预适应小鼠为动物模型，将蛋白和代谢物研究相关联，探究小鼠海马组织与低氧预适应缺氧耐受时间变化相关的蛋白和代谢物，利用蛋白质相互作用数据库、代谢网络数据库以及系统生物学网络建模技术，构建低氧预适应引发的蛋白质表达和代谢调控变化网络，解析低氧预适应神经保护作用的关键节点，初步确认低氧预适应抗缺氧神经损伤的关键蛋白或代谢物，在此基础上在细胞水平上，通过上调或下调这些关键蛋白或代谢物，验证其在低氧适应调控网络中的作用及其机制，以期在分子网络水平阐明低氧预适应抗缺氧神经损伤作用的物质基础及其调控机制，探寻具有自主知识产权的抗缺氧神经损伤关键调节分子和新的药物作用靶标，为发展创新性的抗缺氧神经损伤候选药物提供理论依据。

低氧预适应对缺氧神经损伤的内源性保护作用已有共识，但对其产生的内源性保护作用的物质基础认识仍处于初级阶段。本项目整合iTRAQ蛋白质组学和 UPLC-QTOFMS代谢组学技术，以急性重复低氧预适应小鼠为动物模型，将蛋白和代谢物研究相关联，探究小鼠海马组织与低氧预适应缺氧耐受时间变化相关的蛋白和代谢物，利用蛋白质相互作用数据库、代谢网络数据库以及系统生物学网络建模技术，构建低氧预适应引发的蛋白质表达和代谢调控变化网络。我们的代谢组学结果显示HPC引起海马组织20种代谢物发生显著变化，其中腺苷、5-羟色氨、牛磺酸、γ-氨基丁酸随低氧暴露次数的增加持续升高，而溶血磷脂酰胆碱LPC(16:0)和LPC(18:0)随着低氧暴露次数的增加持续降低，通过MetaboAnalyst 3.0网络数据库进行通路富集分析发现20种代谢物涉及苯丙氨酸、酪氨酸和色氨酸生物合成与代谢调节、甘油磷脂代谢调节。蛋白组学结果显示HPC引起169种蛋白发生变化，主要涉及能量代谢，脂肪酸代谢，缬氨酸、亮氨酸和异亮氨酸代谢，丙氨酸代谢，苯丙氨酸代谢。后续我们将在此基础在细胞水平上，通过上调或下调这些关键蛋白或代谢物，验证其在低氧适应调控网络中的作用及其机制，以期在分子网络水平阐明低氧预适应抗缺氧神经损伤作用的物质基础及其调控机制，探寻具有自主知识产权的抗缺氧神经损伤关键调节分子和新的药物作用靶标，为发展创新性的抗缺氧神经损伤候选药物提供理论依据。