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博碩士論文 etd-0828111-002130 詳細資訊
Title page for etd-0828111-002130
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
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
52
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
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
統計
Statistics
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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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