The mechanism and sex-specific effects of nicotinamide on cue-primed cocaine seeking

项目摘要 目前还没有FDA批准的精神刺激剂使用障碍的药物治疗方法。因此， 开发有效的干预措施，以抵消促使复发的细胞适应 毒品使用仍然是一个优先事项。现有证据表明，使用可卡因会导致氧化标志物升高 应激，在人类和临床前动物成瘾模型中，重要的是，这些标志物可能 助长了寻求毒品的行为。烟酰胺(NAM)是维生素B3的一种，是至关重要的成分 辅酶NAD+、NADP和聚腺苷二磷酸核糖聚合酶(PARP-1)抑制剂可抑制或逆转 氧化应激的标志物。该应用的初步数据表明，系统地给药了NAM 在服用可卡因后，男性自我注射可卡因可以特别地减少恢复到线索引发的寻找，但 不是雌性，是老鼠。这个应用程序的总体目标是测试关于背后机制的假设 NAM减少男性恢复线索的能力。目标1将测试NAM抑制的能力的假设 PARP-1参与了观察到的行为效应。这将通过直接交付 在杏仁中央核(CEA)注射PARP-1抑制剂，然后进行恢复试验(AIM 1a)，作为 以及可卡因自身给药和长期给药后PARP-1活性的测定 (目标1b)。CEA特别令人感兴趣，因为在CEA中直接抑制PARP-1之前 显示减少对可卡因的条件性位置偏爱，CEA已经被明确地牵涉到线索- 已准备好复职。目标2将检验NAM逆转可卡因诱导的特定的 伏隔核(NAC)和CEA中氧化应激的生物标记物。这将通过以下方式实现 自身应激后氧化应激细胞标志物的免疫组织化学和共聚焦成像 管理和灭绝。有趣的是，PARP-1抑制和NAM效应的性别差异 以前有过治疗的报道，但没有药物暴露范例。同样，男性和女性的性别差异 氧化应激是众所周知的，但还没有在药物滥用模型中进行研究。因此， 本提案中概述的实验不仅将告知不结盟运动作为候选干预措施的效用 对药物配对提示的反应，但也将促进我们对基本细胞反应的理解 大脑中含有可卡因。我推测，直接抑制CEA中的PARP-1将减少线索启动 恢复，但不是可卡因引发的恢复，而且不结盟运动管理将减少可卡因诱导的 雄性大鼠PARP-1活性升高(目标1)。此外，我假设不结盟运动政府将减少 雄性大鼠大脑中可卡因依赖的氧化应激，而雌性大鼠则不是(目标2)。总而言之，这 提案将(1)提供定量荧光显微镜方面的培训，(2)将提供关于以下方面的信息 可卡因诱导雄性和雌性大鼠的氧化应激，以及(3)将更清楚地阐明 NAM损害恢复寻求可卡因的机制。

Project Summary No FDA approved pharmacological treatments exist for psychostimulant use disorders. Accordingly, development of effective interventions that can countermand the cellular adaptations which drive relapse to drug use remains a priority. Existing evidence indicates that cocaine use leads to elevated markers of oxidative stress, in both humans and preclinical animal models of addiction, and importantly that these markers may contribute to drug seeking behaviors. Nicotinamide (NAM), a form of vitamin B3, component of crucial coenzymes NAD+ and NADP, and poly(ADP-ribose)polymerase (PARP-1) inhibitor can inhibit or reverse markers of oxidative stress. Preliminary data for this application indicate that systemic administration of NAM following cocaine self-administration can specifically reduce reinstatement to cue-primed seeking in male, but not female, rats. The overall goal of this application is to test hypotheses regarding the mechanism behind NAM’s ability to reduce cue-reinstatement in males. Aim 1 will test the hypothesis that NAM’s ability to inhibit PARP-1 contributes to the observed behavioral effect. This will be accomplished with direct delivery of a PARP-1 inhibitor to the central nucleus of the amygdala (CeA) followed by reinstatement testing (Aim 1a) as well as measurement of PARP-1 activity following cocaine self-administration and chronic NAM administration (Aim 1b). The CeA is of particular interest, as direct inhibition of PARP-1 in the CeA has been previously shown to reduce conditioned place preference to cocaine, and the CeA has been implicated specifically in cue- primed reinstatement. Aim 2 will test the hypothesis that NAM reverses cocaine-induced increases in a specific biomarker for oxidative stress in the nucleus accumbens (NAc) and CeA. This will be accomplished through immunohistochemistry and confocal imaging of a cellular marker for oxidative stress following self- administration and extinction. Interestingly, sex differences in the effects of PARP-1 inhibition and NAM treatment have been previously reported, but not in a drug exposure paradigm. Likewise, sex differences in oxidative stress are well known, but have not been examined in drug abuse models. Therefore, the experiments outlined in this proposal will not only inform the utility of NAM as a candidate intervention for responsiveness to drug-paired cues, but will also advance our understanding of the basic cellular responses to cocaine in the brain. I hypothesize that direct inhibition of PARP-1 in the CeA will reduce cue-primed reinstatement, but not cocaine-primed reinstatement, and that NAM administration will reduce cocaine-induced elevated PARP-1 activity in male rats (Aim 1). Further, I hypothesize that NAM administration will reduce cocaine-dependent oxidative stress in the brains of male, but not female rats (Aim 2). Collectively, this proposal will (1) provide training in quantitative fluorescent microscopy, (2) will provide information regarding induction of oxidative stress by cocaine in male and female rats, and (3) will more clearly elucidate the mechanism by which NAM impairs reinstatement of cocaine seeking.