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論文名稱 Title |
Beta-腦內啡在內皮細胞所造成血管新生的影響 The Angiogenic Effects of β-endorphin in Endothelial Cells |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
52 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2011-07-07 |
繳交日期 Date of Submission |
2011-08-28 |
關鍵字 Keywords |
β-腦內啡、鴉片類受體、嗎啡、血管新生、內皮細胞 angiogenesis, morphine, opioid receptor, endothelial cells, β-EP |
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統計 Statistics |
本論文已被瀏覽 5670 次,被下載 1517 次 The thesis/dissertation has been browsed 5670 times, has been downloaded 1517 times. |
中文摘要 |
血管新生是正常生理中傷口癒合或者在胎兒生長時所必須的過程。過度的血管新生可能導致一些血管新生的疾病,並且被認為是腫瘤生長和轉移所必需的步驟。目前已知pro-opiomelanocortin (POMC)衍生的β-腦內啡(β-endorphin; β-EP)在痛覺、免疫系統方面與減少壓力的調控中扮演著重要角色。但是對於β-腦內啡所引發的血管新生效用仍不清楚。文獻指出嗎啡抑制內皮細胞增生與缺氧誘發之vascular endothelial growth factor (VEGF)釋放。但是,另有文獻發現嗎啡會藉由透過鴉片類受體(opioid receptors)活化Ras,PI3k和MAPK/ERK 的途徑而促使內皮細胞增生。因此,β-腦內啡與鴉片類受體調節血管新生的功能與機轉仍有爭議。為進一步瞭解鴉片類胜肽在血管新生中所扮演的角色與調控機轉,本研究探討β-腦內啡與嗎啡在內皮細胞中所引起的血管新生作用。研究中發現,β-腦內啡有明顯劑量效應的促進內皮細胞增生、移行、與管柱形成。相反的,嗎啡抑制內皮細胞增生、移行、與管柱形成;在離體主動脈環實驗中,β-腦內啡也會促進,而嗎啡卻是產生減少新生微脈管形成。同時發現內皮細胞含有MOR,DOR,KOR等鴉片類受體。利用鴉片類受體拮抗劑Naloxone與μ-鴉片類受體中和抗體能阻斷β-腦內啡與嗎啡對內皮細胞增生、移行之作用。 因此,發現β-腦內啡與嗎啡主要藉由μ-鴉片類受體來正向與負向調控內皮細胞的血管新生作用。因此推測β-腦內啡可能是一種新的促進血管新生的因子。 |
Abstract |
Angiogenesis is a fundamental process in reproduction and wound healing. Angiogenesis is also indispensable for solid tumor growth and metastasis, and also associated with angiogenic diseases. Beta-endorphin (β-EP), derived from its precursor pro-opiomelancortin (POMC), is well known for its role in nociception and immune regulation. However, the function of morphine and β-EP during angiogenesis remains characterization. One previous study indicated that morphine inhibited the proliferation and hypoxia-induced vascular endothelial growth factor (VEGF) release of endothelial cells. Contrastingly, another report found that morphine via Ras/PI3k/MAPK/ERK signaling promotes the survival and angiogenesis in endothelial cells. Besides, endogenous opioid peptides stimulated angiogenesis in chicken allantoic membrane assay through opioid receptors. Thus, the function and mechanism of β-EP and opioid receptors in angiogenesis are controversial. This study evaluated the culture effects of β-EP and morphine on angiogenesis . It was found that β-EP stimulated the proliferation, migration, and tube formation of endothelial cells in a dose-dependent manner. Morphine at a high dose inhibited the proliferation, migration, and tube formation of endothelial cells. In the ex vivo rat aortic ring assay, β-EP enhanced, whereas morphine perturbed, the microvessel sprouting. We also confirmed the expression of MOR,DOR,KOR opioid receptor in endothelial cells. Application of naloxone, a selective opioid antagonist, and neutralizing antibodies of MOR abolished the angiogenic effect of β-EP and morphine. Thus β-EP and morphine exert the pro- and anti-angiogenic effect via MOR, respectively .Besides, β-EP can be regarded as a novel angiogenic factor. |
目次 Table of Contents |
論文審定書--i 中文摘要--ii 英文摘要--iii 目錄 --iv 圖目錄--vi 縮寫檢索表 --vii 第 一 章前言--1 血管新生 --1 血管內皮細胞--2 鴉片類受體--2 嗎啡--3 β-腦內啡--4 第二章研究目的--6 第 三 章 材料與方法--7 細胞培養及試劑--7 細胞繼代培養--7 細胞計數--7 量產及純化腺病毒載體--8 β-EP重組腺病毒--8 細胞增生率測定--9 細胞移行實驗--9 血管管柱形成分析--10 明膠蛋白酵素電泳法--10 一氧化氮生成測定--11 免疫螢光染色--11 蛋白質定量分析--12 SDS-PAGE電泳膠製作--12 西方墨點法分析--12 大鼠主動脈環試驗--13 統計學分析--13 第 四章 實驗結果--14 β-EP促進內皮細胞增生率--14 β-EP促進內皮細胞移行作用--14 β-EP促進內皮細胞血管管柱形成--14 β-EP對 MMP-及 MMP-9釋放影響--15 β-EP於體外主動脈環產生血管新生現象--15 內皮細胞具有鴉片類受體的表現--16 Naloxone抑制β-EP促進內皮細胞增生率與移行的作用--16 β-EP作用在內皮細胞所引發的VEGF/Flk-1,ET-1/ETBR訊息傳遞路徑--17 β-EP影響內皮細胞一氧化氮生成 --17 β-EP作用在內皮細胞引發的Akt/ PTEN/ NFĸB、p38/PI3K訊息 傳遞路徑 --18 第五章 討論--19 第六章 結論-- 22 第七章參考文獻--23 圖表及說明--27 |
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