新型纳米材料用于吗啡成瘾相关的磷酸化蛋白谱分析及分子机制研究

项目名称： 新型纳米材料用于吗啡成瘾相关的磷酸化蛋白谱分析及分子机制研究

项目编号： No.30873132

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 轻工业、手工业

项目作者： 毛煜

作者单位： 中国人民解放军第二军医大学

项目金额： 31万元

中文摘要： 阿片成瘾的分子机制至今尚不清楚。本研究建立了吗啡精神依赖模型(CPP)，同时制备了用于小鼠脑组织磷酸化蛋白富集的纳米材料，对其性能进行表征和优化，并用于吗啡成瘾小鼠脑组织蛋白组学分析。另外通过气相色谱-质谱技术平台，对成瘾相关的各脑区小分子代谢组进行了全面分析，探索在成瘾不同阶段，蛋白和重要信号分子的变化。确证在阿片类药物成瘾过程中起关键作用的分子。通过本项目研究工作，我们发现了吗啡成瘾小鼠脑组织中多个差异表达的磷酸化蛋白。同时通过GC-MS/MS代谢组分析我们还发现Ⅲ#22411;组蛋白乙酰化酶抑制剂在吗啡成瘾小鼠脑组织中含量明显升高。因此，我们认为蛋白的翻译后修饰、转录因子及相关蛋白的抑制与表达、组蛋白修饰的表观遗传学机制等多种分子机制在阿片类药物成瘾过程中扮演了非常重要的作用，其中Ⅲ#22411;组蛋白乙酰化酶可能成为治疗阿片类药物依赖的靶标。

中文关键词： 磁性微球材料；磷酸化蛋白；阿片成瘾；组蛋白；分子机制

英文摘要： The mechanisms that underlie the development of opioid addiction still remain unclear. In this study, morphine-induced conditioned place preference (CPP) in mice was used as an animal model to explore the mechanism of context-dependent learning associated with rewarding effect of morphine. We also initiated a new material named Fe3+ -immobilized magnetic silica microsphere, which can efficiently and selectively enrich trace phosphopeptides with several hundred times of enrichment efficiency. The enriched peptides were then identified by high sensitive MALDI-TOF Mass Spectrometry platform. The differntially expressed phosphopeptides associated with opioid addiction were identified by this paltform. At the same time the metabolite profiles of brain tissues of morphine-treated and saline-treated mice were compared using GC-MS/MS. Nicotinamide, as a competitive inhibitor of the class III NAD+-dependent HDACs, was found increased for CPP mice evoked by morphine treatment. According to our work, several combination of mechanisms may underlay drug abuse, which including protein post-translational modification, transcription factor and relative targeted proteins, histone modification. The class ⅢNAD+-dependent HDACs can be an therapy targets of opioid addiction.

英文关键词： Magnetic silica microspheres; Protein phosphorylation;Opioid addiction; histone；Molecular mechanism