2026-11-092026-11-09https://scholars.csmu.edu.tw/handle/123456789/38217Amphetamine (AMPH) and associated derivatives are the most commonly abused psychostimulant in the young people. AMPH is known for its suppressive effect on appetite. A daily treatment of AMPH for several days could induce a marked decrease of food intake on the first day of dosing and then the food intake returned gradually to the normal level (tolerant effect) on the followings. The mechanism for the anorectic action of AMPH was through the inhibitory action on hypothalamic neuropeptide Y, an orexigenic neurotransmitter in the brain. Amphetamine is an artificially synthesized psychostimulant. It could activate both central and autonomic nervous system. As we know, amphetamine acts on mesolimbic circuit or associated brain regions and influences on associated emotional circuit. Therefore, to investigate the modulation of amphetamine on emotional circuit is very important for current study. Cocaine and Amphetamine Regulated Transcript (CART), brilliantly identified by Douglass and co-workers is an anorectic peptide regulated by leptin. Fasting animals show a pronounced decrease in CART mRNA expression in the arcuate nucleus, an area of the brain known to be associated with feeding behavior. In animal models with disrupted leptin signalling, CART mRNA is almost absent in the arcuate nucleus. ICV administration of CART inhibits both normal and starvation-induced feeding and completely blocks the feeding response induced by NPY. Antisera directed against CART increases feeding in normal rats. Indications are that CART may be an endogenous inhibitor of food intake Therefore, the following objectives will be done in this project, including (1) to examine the effect of repeated AMPH treatments on feeding behavior, rats were injected intraperitoneally (i.p.) with the drug at doses of 0, 2 or 4 mg/kg daily for 4 days. The first time to inject AMPH was at the end of Day 0. The intake data was calculated with respect to the food amount of previous day. (2) to investigate the impacts of AMPH on the expression of genes, such as CART,SOD, ROS and NOS via western blotting and to further analyze their mRNA levels via RT-PCR assay. (3) Using CART antisense oligodeoxynucleotide to investigate the mechanism in AMPH. (4) investigate the regulation of transcription factors via western blotting and EMSA assay to determine the transactivity of these transcription factors such as, NF-kB, c-Jun, c-fos.自從安非他命 (Amphetamine)被濫用以來，社會各界普遍認為藥物濫用日益嚴重，而衛生署管制藥品管理局也發現台北地區非法用藥盛行率為1.5％，非法用藥種類之前三名為安非他命、大麻、強力膠。因此，除了如何杜絕安非他命的濫用以外，安非他命的毒性機轉探討也是目前很重要的課題之ㄧ。 安非他命對於中樞神經系統的作用，大都來自對於諸多神經傳導物質受體系統的影 響，經由在貯存處游離釋放、阻斷回收、抑制代謝的方式，增加神經傳導物質的作用強度和時間，造成中樞神經的興奮、神經毒性和行為的變化。而目前已有許多研究證實安非他命對於norepinephrine 受體系統的作用，會導致中樞神經興奮；而對於dopamine 受體系統的作用則是產生快感和幻覺，並強化回饋成癮系統 (reward system) ；至於對於serotonin 的回收抑制則先是造成分泌增加，進而因回饋機制導致接受器的敏感度降低，造成serotonin急速消耗，這時候使用者會出現憂鬱、焦慮、注意力不集中、記憶力障礙和失眠問題，而長期使用則會導致腦功能受損和神經毒性。 CART的全名為Cocaine and Amphetamine-Regulated Transcript (古柯鹼和安非他命調控之轉錄因子)。目前研究顯示CART peptide與Amphetamine或古柯鹼等興奮性濫用藥物之作用有關，也有許多的文獻指出CART 55-102是調控著厭食反應最重要的區域。這結果顯示CART可能是一種內生性的神經傳遞物質，可能會影響許多與厭食反應有關的訊息傳遞，但其詳細機制及其相關的訊息傳遞路徑卻仍不清楚。 雖然安非他命對中樞神經的影響已有許多相關的研究，但對於下視丘及其他腦區的影響及對於CART的影響則仍不清楚，因此本計畫擬 (1)利用大白鼠以「重複注射Amphetamine」的處理方式，每天一次，連續數天。在重複注射期間，每天記錄大白鼠的食物攝取量，完成由Amphetamine誘發之毒性及厭食的影響。(2)利用同樣的模式在重複注射的不同時間犧牲大白鼠，取腦內的下視丘及其他腦區，萃取其蛋白及mRNA，以western blotting、RT-PCR及Real time RT-PCR的分析方式，完成不同時間Amphetamine對下視丘及其他腦區中CART、SOD、ROS及NOS的基因變化及影響。(3)另外，我們擬設計CART的antisense oligodeoxynucleotide，以活體(In Vivo)的動物為材料，在Amphetamine注射前，將CART的antisense oligodeoxynucleotide直接注入下視丘PVN部位，由攝食行為的改變及CART基因的表現，觀察改變CART的antisense對Amphetamine之影響。並進一步觀察在CART antisense的注射下，大白鼠下視丘及其他腦區中SOD、ROS及NOS的基因變化及影響。(4) 最後，我們擬利用western blotting及EMSA的分析方式，探討經由Amphetamine處理大白鼠其下視丘及其他腦區一些常見的transcription factor如NF-kB、c-Jun、c-fos及JNK其蛋白質表現及其與相關DNA結合能力的變化。安非他命古柯鹼及安非他命調節轉錄因子訊息傳遞AmphetamineCARTsignal transductionProject