Neuropathic mechanisms and gene therapy on opioid dependence

描述（由申请人提供）：阿片类药物依赖是一种慢性和复发性疾病，以强迫性药物寻求和使用为特征。阿片类药物依赖导致过度的身体依赖（戒断综合征）。虽然对阿片类药物的心理依赖是依赖/滥用的重要特征，但身体依赖本身（戒断综合征）是强迫性吸毒行为和短期复发的主要原因。遗憾的是，阿片类药物在阿片类药物依赖病理改变中的作用机制尚不清楚。最近的研究表明，慢性吗啡可诱导促炎细胞因子（如TNFα）的释放。氧化应激可能参与海洛因依赖/戒断过程。然而，很少有研究解决这些胶质细胞因子在阿片类药物戒断中参与中枢神经系统TNFR、钙和ROS反应的确切机制。PAG在阿片类药物戒断中的作用已被研究提出，但我们对其详细的分子机制知之甚少。在这里，我们假设神经胶质激活导致促炎细胞因子的释放，细胞因子受体激活的神经元诱导线粒体ROS和pCREB（药物滥用行为的关键蛋白）在吗啡戒断的发展中。1、研究吗啡戒断时神经胶质TLR4激活是否诱导细胞因子的释放。目的2、观察细胞因子受体的激活是否诱导吗啡戒断大鼠PAG线粒体ROS的发生。特异性目的3、检测PAG线粒体ROS是否通过pCREB参与MW。该提案将使用药理学，分子生物学，免疫组织化学，细胞类型靶向基因转染和成像密度分析的分析来完成新的假设。这些研究结果将为阿片类药物滥用的发病机制提供重要见解，并可能为阿片类药物依赖的新疗法提供临床前证据。

DESCRIPTION (provided by applicant): Opioid dependence is a chronic and relapsing disease and characterized by compulsive drug seeking and use. Opioid dependence results in excessive degrees of physical dependence (withdrawal syndrome). Although psychological dependence on opioids is an important feature of dependence/abuse, physical dependence itself (withdrawal syndrome) is a major cause of compulsive drug-taking behavior and short term relapse. Unfortunately, the mechanism of opioid in the pathological changes in opioid dependence is not yet understood in detail. Recent studies show that chronic morphine induces release of proinflammatory cytokines (e.g. TNFα). Oxidative stress may participate in the process of dependence /withdrawal of heroin. However, very few studies have addressed the precise mechanisms by which these glial elements contribute to CNS neuronal TNFR, calcium and ROS response in opioid withdrawal. The role of the PAG in the expression of opioid withdrawal has been suggested by studies, but we know little of the detailed molecular mechanisms. Here, we hypothesize that glial activation resulting in the release of proinflammatory cytokines, and that neurons activated by cytokine receptor induce mitochondrial ROS and pCREB (a key protein in drug abuse behavior) in the development of morphine withdrawal. Specific Aim 1, To test whether glial TLR4 activation induces release of cytokine in morphine withdrawal. Specific Aim 2, To test whether activation of cytokine receptor induces mitochondrial ROS in morphine withdrawal in the PAG. Specific Aim 3, To test whether mitochondrial ROS in the PAG is involved in MW through the pCREB. The proposal will use assays of pharmacology, molecular biology, immunohistochemistry, cell type-targeting gene transfection, and imaging density analysis to complete the novel hypothesis. The results of these studies will provide important insights into the pathogenesis of opioid abuse, and may provide preclinical evidence for a novel therapy of opioid dependence.