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系統識別號 U0026-2508201400050800
論文名稱(中文) 半乳糖凝集素-1在慢性阻塞性肺病的病理生理學角色
論文名稱(英文) Role of galectin-1 in the pathogenesis of chronic obstructive pulmonary disease
校院名稱 成功大學
系所名稱(中) 微生物及免疫學研究所
系所名稱(英) Department of Microbiology & Immunology
學年度 102
學期 2
出版年 103
研究生(中文) 王崇騰
研究生(英文) Chung-Teng Wang
學號 S46011029
學位類別 碩士
語文別 中文
論文頁數 48頁
口試委員 指導教授-蕭璦莉
口試委員-吳昭良
口試委員-曾堯麟
口試委員-陳玉玲
口試委員-張孟雅
中文關鍵字 半乳糖凝集素-1;前胸腺素  肺氣腫  菸萃取液 
英文關鍵字 galectin-1  chronic obstructive pulmonary disease 
學科別分類
中文摘要 肺氣腫 (Emphysema) 是主要的一種慢性阻塞性肺病 (Chronic Obstructive Pulmonary Disease, COPD) 的典型,病徵為肺泡壁的瓦解以及肺泡氣室的異常擴張,導致肺臟失去彈性,呼吸系統失能。根據世界衛生組織估計,慢性阻塞性肺疾病將於 2020 年成為全球人類的第三大死因,對人類健康的影響甚鉅。引起肺氣腫的主因為長期性的抽菸,然而卻僅有 20% 的抽菸族群成為肺氣腫患者,其中顯然有未定義的因子參與。前胸腺素 (Prothymosin α, 簡稱 ProT) 在免疫與氧化壓力的調節扮演重要角色,在過去實驗室的研究發現 ProT 過量表現的同型合子小鼠會自發性的產生肺氣腫,並且在肺氣腫患者的組織切片中看到 ProT 有過量表現。半乳糖凝集素-1 (galectin-1)屬於S型凝集素,galectin-1在生物體中的功能相當複雜,包含調控細胞間連結、發炎調控和細胞凋亡過程等。因此,本研究主要探討galectin-1在菸萃取物引起的肺氣腫模式之間的關聯。我們利用galectin-1基因剔除鼠做香菸誘導肺氣腫模式的研究,發現H& E染色的肺切片顯示 galectin-1基因剔除鼠較野生型小鼠肺泡空間擴大和肺泡壁的的破壞更為嚴重,並且galectin-1基因剔除鼠與野生型小鼠相比免疫細胞浸潤以及凋亡細胞明顯增加;並且我們發現給予菸萃取液的小鼠在肺泡及氣管區域會表現galectin-1,但是在 轉殖基因鼠的則發現galectin-1表現量相較於野生型小鼠受到抑制。細胞實驗也發現外給galectin-1蛋白能藉由產生細胞自噬現象,清除受損粒線體,幫助肺癌上皮細胞A549抵禦氧化壓力,顯著減少氧化壓力造成的細胞死亡。綜合以上結果,本篇研究指出galectin-1在香菸誘導的肺氣腫模式中扮演保護性角色。
英文摘要 Our researches focus on investigating the association between galectin-1 and chronic obstructive pulmonary disease (COPD), and the main findings include: 1. galectin-1 may play a protective role in CS-induced COPD through its anti-inflammatory activity, 2. prothymosin α suppresses galectin-1 expression, 3. galectin-1-induced autophagy attenuates ROS-induced cell apoptosis. Collectively, our studies suggest that galectin-1 may play a protective role in CS-induced COPD through anti-inflammatory and anti-apoptotic activities. Our results also reveal that ProT may down-regulate galectin-1 expression and thus lead to COPD, especially upon CS exposure.


論文目次 中文摘要. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .I
Abstract. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .III
致謝. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .VIII
總目錄. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .IX
圖目錄. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . XII
縮寫. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .XIII
第一章 緒論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
一、 慢性阻塞性肺疾病(COPD)與吸菸 . . . . . . . . . . . . . . . . . . . ….. . 1
二、 半乳糖凝集素-1 (galectin-1, Gal-1) . . . . . . . . . . . . . . . .. . . … 3
三、 前胸腺素 (Prothymosin α, ProT). . . . . . . . . . . . . . . . . . . . . .4
第二章 研究目的. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6
第三章 材料與方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
一、材料.. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
1、 菸萃取液. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
2、 實驗動物. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
3、 細胞株. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........8
4、 引子. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
5、 質體. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .8
6、 試劑. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9
7、 勝任細胞. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11
8、 抗體. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .11
二、 方法. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
1、 菸萃取液誘導肺氣腫之動物模式. . . . . . . . . . . . . . . . . . . . .. .. .13
2、 免疫組織化學染色法 (IHC). . . . . . . . . . . . . . . . . . . . . . . . . .14
3、 肺泡擴大的百分比. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .14
4、 肺氣腫發病率. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15
5、 慢病毒生產. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .15
6、 galectin-1蛋白的生產. . . . . . . . . . . . . . . . . . . . . . . . . . . .16
7、 galectin-1蛋白的純化 . . . . . . . . . . . . . . . . . . . . . . . . . . .16
8、 濃縮重組galectin-1蛋白. . . . . . . . . . . . . . . . . . . . . . . . . . .17
9、 細胞培養. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18
10、 RNA萃取. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .18
11、 反轉錄酶聚合連鎖反應 (RT-PCR). . . . . . . . . . . . . . . . . . . . . . .19
12、 即時半反應反轉錄酶聚合連鎖反應 (Real-time PCR) . . . . .. .. .. .. .. .. .19
13、 細胞氧化壓力存活率測試. . . . . . . . . . . . . . . . . . . . . . . . . ..20
14、 西方墨點法 (Western blot). . . . . . . . . . . . . . . . . . . . . . . . .21
第四章 結果. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
一、Gal-1 基因剔除鼠對於菸誘導的肺氣腫模式具較高感受性. . . . .. . . . .. . . .22
二、前胸腺素(ProT)過量表現抑制在Gal-1表現. . . . . . . . . . . .. . .. . . . . 23
三、Gal-1減少氧化壓力引起的細胞死亡. . . . . . . . .. . . . . . . . . . . . . .24
四、Gal-1 引起細胞自噬現象減少粒線體損壞 . . . . . .. . . . . . . . . . . . . .25
第五章 結論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26
第六章 討論. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
參考文獻. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .31
圖表. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .37
附錄. . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . .. . . .48
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