2010-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649316摘要：憂鬱症是精神疾病中盛行率極高的情感性疾病，世界衛生組織以DALY為指標，估算出憂鬱症將在2020年成為排名第二的高負擔疾病。由於目治療方式會帶來許多病人無法接受的副作用，因此尋找替代治療方法是具有發展潛力的課題。天麻是中醫食療中被用作養心安神的材料，本研究室先前的研究也發現天麻水萃物在動物實驗中具有顯著的抗憂鬱效果，同時能夠增加腦中血清素與多巴胺的含量、同時降低血清素與多巴胺的代謝速率，然而其更詳細的作用機制尚未明瞭。因此本實驗擬以細胞實驗、以及基因體學與蛋白質體學的研究，來探討天麻水萃物的抗憂鬱機制。本計畫擬分三年完成： 第一年：以MTT assay與免疫染色法來瞭解不同濃度天麻水萃物對細胞生存力與形態之影響。隨後 進行單胺氧化&#37238;活性之試驗，與神經保護之細胞試驗，以瞭解天麻水萃物是否具有抑制單胺氧化&#37238;活性或保護細胞免於corticosterone之損傷之能力。 第二年：自基因的層次來探討天麻可能的作用機制。以cDNA microarray之技術來偵測大鼠腦組織之基因表現，藉此尋找出天麻可能的作用基因，以及可能的抗憂鬱基因。 第三年：以第二年研究中得到的基因作為目標基因，以二維電泳分析，並以MALTI-TOF MS來比較其腦組織蛋白質表現量的差異。期望可以比對出最可能的作用蛋白質。<br> Abstract: Depression is one of the most common psychiatric disorders with high mortality and morbidity. The medication used in clinics nowadays brings lot of side effects. Thus, looking for an alternative therapy for depression is an important work. Gastrodia elata Bl. is a Chinese medicine used for centuries. It was found that the water extract of Gastrodia elata Bl. (WGE) possesses antidepressant effect in animal mode.Serotonin and dopamine content was increased, and the turnover of serotonin and dopamine were decreased after WGE administration. But the antidepressant mechanism of Gastrodia elata Bl. is unknown. Therefore, the present study will investigate the antidepressant mechanisms of WGE via cell model and genomics and proteomics study. This project will be performed in three years: First year: The cell viability and cell morphology will be studied by MTT assay and immunostaining respectively to investigate the effects of different concentrations of WGE on PC12 cell line. The monoamine oxidase activity and the cell viability and the intracellular calcium concentration after corticosterone treatment will be studied for evaluating the effect of WGE on the monoamine oxidase activity and the neuroprotective activity. Second year: The genes will be focused in this part of study. The possible genes expression related to WGE treatment will be detected by microarray. And the QRT-PCR was performed to confirm the gene expression changes observed by microarrays. Third year: The altered genes found in the 2nd year will be regarded as target genes. Protein extracts from rats’ brain, including frontal cortex, hippoacmpus, amygdala, and striatum, were processed for 2D gel characterization. Maps were analyzed by the PDQuest image analysis software (Bio-Rad). And the differentially expressed proteins were identified by MALDI-TOF mass spectrometry.天麻憂鬱單胺氧化&#37238基因體學蛋白質體學Gastrodia elata Bl.depressionmonoamine oxidasegenomicsproteomics