2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38807Neuropeptide Y (NPY) is an orexigenic agent abundant in the brain, while POMC (proopiomelanocortin) is an anorexigenic neurotransmitter. The mechanism for the anorectic effect of amphetamine (AMPH) is linked to the central release of catecholamine, which in turn inhibits the expression of NPY gene. Daily treatment with AMPH for several days induced a marked anorexia (AMPH anorexia) on day 1 and day 2, which followed by a gradual restoration of feeding behavior (AMPH tolerance). The response of AMPH anorexia is related to the inhibition of NPY gene but the response of AMPH tolerance is relevant to the restoration of NPY gene activity. Recently, from the study exploring the possible role of PKA (or PKC) signaling in the regulation of NPY gene, we found that POMC gene might involve in the action of catecholamine on NPY neurons. Thus, the present study is aimed to assess this hypothesis. The proposal for the present investigation can be divided into three parts (Part 1-Part 3) that will be prosecuted within 3 consecutive years. The aim of Part 1 is to investigate the role of POMC in the regulation of NPY gene expression and AMPH-induced appetite suppression. The aim of Part 2 is to examine the role of POMC gene in the regulation of anti-oxidative enzyme, i.e., superoxide dismutase (SOD) or nitric oxide synthase (NOS), when the animal is in conditions of AMPH neurotoxicity. The aim of Part 3 is to examine the role of signal transduction (PKA, PKC, CREB, AP-1, etc.) in the regulation of POMC-mediated anorexia during AMPH treatment. There are three major experiments that need to be executed for each Part of investigation. In Part 1, it included experiments that examine: (1) the effect of AMPH treatment on POMC and NPY gene expressions, (2) the effect of AMPH treatment on hypothalamic POMC and NPY immunoreactivity by double-immunoblotting technique, and (3) the effect of POMC knock down in the brain on NPY gene expression and AMPH-induced anorexia. In Part 2, it included experiments that examine: (1) the effect of AMPH neurotoxicity on POMC, SOD, NOS and NPY gene expression, (2) the effect of POMC knock down in the brain on AMPH neurotoxicity, and (3) the effect of free radical inhibitor on POMC gene expression. In Part 3, it included experiments that examine: (1) the effect of AMPH treatment on POMC, PKA, PKC, CREB, AP-1 and NPY gene expression, (2) the effect of AMPH treatment on POMC, SOD and NOD immunoreactivity by double-immunobloting technique, and (3) the effect of PKA (or PKC, CREB, AP-1, etc.) knock down on POMC-mediated anorexia. Moreover, in each Part of the investigation, there are two sets of animals, i.e., AMPH anorexia and AMPH tolerance groups, which will be used to study the acute and chronic treatment of AMPH, respectively. After the complete examination of AMPH, we will further investigate the effect of its analogue, i.e., amphetamine-like anorectic drugs.Y 型神經胜肽(neuropeptide Y, NPY)是腦內含量豐富可興奮食慾的神經傳遞物，而原始類 鴉片黑色素皮質素(proopiomelanocortin, POMC)是腦內厭食性神經傳遞物。本文所指厭 食劑是指安非他命(amphetamine, AMPH)及其類似物。安非他命的厭食作用，是透過腦 內catecholamine 的釋放，去抑制腦內NPY 基因所致。安非他命若每天注射一次，持續 數天時，在第一天及第二天可引發最顯著的厭食效應(本文稱為AMPH anorexia)，但在 隨後數天，則逐漸產生厭食耐受性(本文稱為AMPH tolerance)。AMPH anorexia 與「抑 制NPY 基因的表現」有關，而AMPH tolerance 則與「NPY 基因表現的逐漸回升」有關。 最近，後學由蛋白激酶(PKA、PKC)等訊息傳遞實驗發現，安非他命對NPY 的抑制作用 並非直接的，可能透過CART(cocaine/AMPH-regulated transcript)路徑或POMC 路徑媒介 此厭食作用；然而，也發現CART 路徑只是扮演微調控角色，或許POMC 路徑才是主 要調控角色。因此，本計畫將探討POMC 路徑參與之可能性。計畫名稱定為：下視丘 POMC 神經傳遞路徑在類安非他命藥物引發的生化反應及厭食作用所扮演之角色。生化 反應包括NPY 基因表現、毒性及訊息傳遞。擬分三年探討，第一年子計畫名稱為：「下 視丘中POMC 神經路徑在類安非他命厭食劑作用之角色」，第二年名稱為：「POMC 在 安非他命誘發之毒性之角色」,第三年名稱為：「訊息傳遞在POMC 媒介之厭食效應之 角色」。每個子計畫的實驗將分「AMPH anorexia」及「AMPH tolerance」兩組動物，分 別探討急性及慢性安非他命的作用。第一年預期完成三項工作：(1)安非他命注射對 POMC 基因表現的影響，(2)以「雙重免疫顆粒染色法」觀察POMC 及NPY 免疫顆粒之 表現，(3)腦內POMC 基因剃除(knock down)對NPY 基因及厭食行為的影響。第二年預 期完成三項工作：(1)安非他命中毒(AMPH neurotoxicity)對POMC 基因, 及抗氧化基因， 例如superoxide dismutase (SOD), nitric oxide synthase (NOS)等基因表現的影響，(2)腦內 POMC 基因剃除對安非他命中毒的影響，(3)自由基抑制劑 (melatonin 或7-nitroindazole) 前處理對POMC 基因表現的影響。第三年預期完成三項工作：(1)安非他命對POMC、 PKA、PKC、CREB、c-fos 及c-jun 基因表現的影響，(2)以「雙重免疫顆粒染色法」觀 察POMC 及PKA (or PKC 等)免疫顆粒之表現，(3)下視丘旁室核剃除PKA (或PKC、 CREB、AP-1)基因，對POMC 路徑及厭食行為的影響。以上安非他命之研究完成後， 將繼續其類似物的研究。原始類鴉片黑色素皮質素(POMC)安非他命中毒攝食行為Y 型神經胜肽(NPY)訊息傳遞蛋白激酶A (PKA)cAMP 反應因子結合蛋白(CREB)抗氧化基因組織免疫化學POMC (proopiomelanocortin)amphetamine neurotoxicityneuropeptide Y(NPY)signal transductionfood intakefree radicalgene knock downimmunohistochemistryoxidative stressProject