系統識別號 U0026-0812200911431775
論文名稱(中文) 人類腹主動脈瘤血管平滑肌細胞之反應性氧化產物的生成及NAD(P)H氧化酶的表現
論文名稱(英文) ROS production and the NADPH oxidase expression in human abdominal aortic aneurysm-derived smooth muscle cells
校院名稱 成功大學
系所名稱(中) 細胞生物及解剖學研究所
系所名稱(英) Institute of Cell Biology and Anatomy
學年度 93
學期 2
出版年 94
研究生(中文) 陳應輝
研究生(英文) Ying-Hui Chen
電子信箱 jenjer.chan@msa.hinet.net
學號 t9692101
學位類別 碩士
語文別 英文
論文頁數 47頁
口試委員 口試委員-陳麗玉
中文關鍵字 反應性氧化產物  腹主動脈瘤  血管平滑肌細胞 
英文關鍵字 abdominal aortic aneurysm  vascular smooth muscle cell  ROS 
中文摘要   在美國超過六十歲的成年人當中有3-8%患有腹主動脈瘤,腹主動脈瘤好發於腎動脈以下至腸動脈之間的腹主動脈,動脈瘤會漸漸擴張或像汽球狀般腫大,引發此疾病的因素包括有細菌或病毒感染,先天性血管結締組織疾病或由外部創傷所引起,這疾病除了會無預警發生動脈瘤破裂外,通常是沒有臨床症狀而成為一個潛伏的疾病,大部份的腹主動脈瘤會伴隨著高血壓及動脈硬化的症狀,雖然真正引起此疾病的原因仍未完全了解,但在最近的十年中反應性氧化產物(ROS)的參與被認為會引起腹主動脈瘤的發生, ROS是氧經氧化成水的過程中所產生一些有害的代謝性產物,而NAD(P)H氧化酶被認為在大部份的血管疾病中會產生大量的ROS,Miller等人曾報告動脈瘤所產生的ROS比鄰近的組織多,在本實驗中,我們假設由人類腹主動脈瘤所培育出來的主動脈平滑肌細(HASMC)胞會產生更多的ROS併伴隨著更强的NAD(P)H氧化酶活性,本實驗共收集了六位病人接受腹主動脈瘤手術的檢體作研究,而以接受冠狀動脉繞道手術的病人,手術過程中在升主動脈打洞所取得的檢體作為對照,本實驗以伸展法培育以取得所需細胞,並經由特定標記的免疫螢光染色證實培育出來的細胞確定為HASMC,實驗組及對照組的HASMC在培養中有相似的形態表現及螢光染色,MTT法測得的細胞增殖速率在兩組間沒有差異,以血管張力素II刺激後經由Dihydroethidium促成螢光產生的實驗中,實驗組比對照組有更强的螢光表現,若在未經血管張力素II刺激下,螢組的基礎值並沒有不一樣,在NAD(P)H氧化酶活性的實驗中,經血管張力素II刺激後實驗組比對照組有兩倍的基礎值上升量,由以上這些實驗可以更明確指出ROS及NAD(P)H氧化酶在人類腹主動脈瘤扮演著重要的角色.
英文摘要  Abdominal aortic aneurysms (AAAs) are present in 3-8% of people over 60 years of age in America. It usually occurs below the renal arteries and may extend into the iliac arteries. Progression of AAA involves dilation, stretching, or ballooning of the aorta. The causes of AAA may include infection, congenital weakening of the connective tissue component of the artery wall, or trauma. This is often a silent disorder until the catastrophic event of aneurysm rupture occurs. Most AAAs occurs in association with advanced atherosclerosis and hypertension. In the past decade, reactive oxygen species (ROS) have been proposed to contribute to the pathogenesis of aneurysm though the exact cause is still unknown. ROS is often considered cytotoxic metabolites during oxidative process from oxygen to water. NAD(P)H oxidase has been reported as a major enzyme for producing ROS in most of the vascular inflammatory diseases. Miller et al showed that tissue of aneurysm produced more ROS then adjacent area. In this study, we hypothesized that AAA-derived human aortic smooth muscle cells (HASMCs) are capable of more ROS production accompanied by higher NAD(P)H oxidase activity. Six AAA specimens were collected from surgery under normal surgical procedure. Specimens from the punctured hole of ascending aorta in patients with coronary artery bypass graft surgery were collected as the controls. HASMCs were cultured from medial layer of the specimens by explant method and the identity of HASMCs was verified by immunostaining with specific markers. AAA-derived and control HASMCs exhibited similar morphological and immunostaining features in culture. The proliferation rate determined by MTT assay showed no difference between the two groups. Angiotensin II-stimulated superoxide anion production as detected by dihydroethidium-derived fluorescence was markedly greater in AAA-derived HASMCs compared to the control HASMCs while no difference was detected in the absence of stimulation. Similarly, angiotensin II stimulation resulted in 2-fold in NAD(P)H oxidase activity assay in AAA-derived HASMCs compared to control HASMCs with similar basal activity in both groups. The above experiments may characterize the important roles of ROS and NAD(P)H oxidase in mediating the disease of AAA.
論文目次 Chinese Abstract……………………………i
Table of contents…………………………vi
Materials and Methods……………………12
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