博碩士論文 101331013 詳細資訊




以作者查詢圖書館館藏 以作者查詢臺灣博碩士 以作者查詢全國書目 勘誤回報 、線上人數:37 、訪客IP:54.208.73.179
姓名 李啟全(LEE, CHI-CHUAN)  查詢紙本館藏   畢業系所 生物醫學工程研究所
論文名稱 以板式流道系統模擬血管內皮細胞於層流剪力影響下受尼古丁刺激產生發炎反應之研究
相關論文
★ 研究探討層流剪應力於高糖環境下對膀胱癌細胞遷移與侵襲行為之影響★ 研究探討層流剪應力對泌尿上皮細胞癌於細胞週期運作之影響與機轉
★ 設計並建構一全氟碳光生物反應器組用於分離混合氣體中之二氧化碳並同時提升微藻養殖及其經濟產物生成之效能★ Synthesis, Spectral Characterization and Evaluation of Quercetin-Zinc Complex for Tumoricidal and Anti-metastasis of Human Bladder Cancer Cell
★ 包覆靛氰綠與喜樹鹼之標靶全氟碳奈米乳劑 研製於強化乳癌螢光擴散光學影像暨 光/化學治療之研究★ 研製包覆靛氰綠與絲裂黴素C之標靶全氟碳奈米乳劑應用於膀胱癌光-化學治療之研究
★ 研製包覆靛氰綠及利福平之聚乳酸-聚甘醇酸奈米粒子用於破壞生物膜之抗菌治療★ 可動態改變外翻力矩的治療退化性膝關節炎輔具
★ 聚乙二醇對於擬球藻生長與脂質堆積之影響★ 製備包覆靛氰綠及阿黴素之聚乳酸甘醇酸-聚乙二醇交聯標靶奈米粒子用於乳癌光/化學治療之研究
★ 研製包覆靛氰綠與阿黴素之標靶氟化奈米乳劑用於乳癌光/化學治療之研究★ 研究設計全氟碳化物光生物反應器系統用以純化沼氣並藉此提升微藻生物質及生質能源之產量
★ 針對糖尿病足潰瘍設計並製作一種抗菌且能促進傷口癒合的甲殼素複合式水凝膠之研究★ 利用PLGA微球載體結合超聲波駐波場以提高巨噬細胞藥物輸送之效率
★ 以血流動力系統探討血管內皮細胞在尼古丁刺激下對層流剪應力之型態異常與自體凋亡之表現變化★ 結合超聲波駐波場與層堆疊自體組裝微球載體建構提高分子傳遞至細胞內效率之方法
檔案 [Endnote RIS 格式]    [Bibtex 格式]    [相關文章]   [文章引用]   [完整記錄]   [館藏目錄]   [檢視]  [下載]
  1. 本電子論文使用權限為同意立即開放。
  2. 已達開放權限電子全文僅授權使用者為學術研究之目的,進行個人非營利性質之檢索、閱讀、列印。
  3. 請遵守中華民國著作權法之相關規定,切勿任意重製、散佈、改作、轉貼、播送,以免觸法。

摘要(中) 尼古丁已被廣泛證明為心血管病變如動脈粥樣硬化(Atherosclerosis)主要的危險因素之一。然而,在先前的研究中缺少了一項影響人類血管內皮生理的重要因子:層流剪應力(Laminar Shear Stress),因此尼古丁在實際血管內如何導致內皮細胞傷害的機制是有待證實的。在本研究中,我們利用人類臍靜脈內皮細胞(Human umbilical vein endothelial cells; HUVECs)作為實驗細胞,施以尼古丁做藥物處理,並結合平行板流道系統模擬層流剪應力,進而來探討血流動力學在尼古丁介導的人類內皮細胞發炎情況的影響。首先HUVECs會以12 dynes/cm2在±10-4M尼古丁的環境下以不同的時間沖刷:時間設定為0(靜態組), 6, 12, 24 小時,接著以血球貼附實驗、活性氧自由基測量,以及西方墨點法分析細胞貼附因子表現等方法來說明內皮細胞的發炎情形。實驗結果顯示,HUVECs在沒有尼古丁的環境下,層流剪應力在24小時內對血球貼附率沒有顯著影響(P > 0.05);而HUVECs在沒有層流剪應力(靜態)的環境下,尼古丁在12小時以及24小時分別提升了血球貼附率1.8倍(P < 0.05)以及2.6倍(P < 0.05)。然而,在結合尼古丁以及層流剪應力處理24小時的組別中,血球貼附率會比以單獨尼古丁處理24小時的組別高1.4倍(P < 0.05)。而此結果與活性氧自由基測量的結果是呈現一致的;在結合尼古丁以及層流剪應力處理24小時的組別中,細胞產生的活性氧會分別比以單獨尼古丁處理24小時或單獨以層流剪應力處理24小時的組別提升2.55倍(P < 0.05)以及4.22倍(P < 0.05)。接著我們以Intercellular Adhesion Molecule-1 (ICAM-1)以及Vascular Adhesion Molecule-1 (VCAM-1)兩種貼附因子蛋白作西方墨點法實驗,分析內皮細胞在尼古丁以及層流剪應力處理後的表現,我們發現ICAM-1以及VCAM-1的表現在單獨尼古丁處理24小時的情況下會分別提升1.3倍(P < 0.05)以及1.9倍(P < 0.05);而在單獨以層流剪應力處理24小時的情況下ICAM-1以及VCAM-1的表現並無顯著的提升(P > 0.05);然而,尼古丁結合層流剪應力則會使ICAM-1表現比單獨尼古丁處理情況下提升1.6倍(P < 0.05),不過卻會使VCAM-1表現下降0.9倍(P > 0.05),此結果顯示了在有尼古丁環境下,層流剪應力在調節內皮細胞上ICAM-1以及VCAM-1的表現扮演了不一樣的角色。本研究顯示了內皮細胞在同時受到尼古丁與層流剪應力刺激24小時的發炎程度會比單獨受到尼古丁或是層流剪應力刺激有顯著的提升(P值均小於0.05),此一結果顯示了層流剪應力在尼古丁造成的內皮細胞發炎反應上扮演了一個不利的角色。
摘要(英) Nicotine has been widely demonstrated as one of major risk factors for cardiovascular lesions such as atherosclerosis. However, the definite mechanism of how nicotine causes endothelial dysfunction remains unclear due to lack of hemodynamic factor in most of prior in vitro studies. In this study, we aimed to investigate the effectiveness of hemodynamic stress on nicotine-mediated human endothelial inflammation using human umbilical vein endothelial cells (HUVECs) and parallel-plate flow system to simulated blood flow-induced laminar shear stress (LSS). HUVECs were first sheared by 12 dynes/cm2 using medium with and without 10-4 M nicotine for 0 (static culture), 6, 12, and 24 h, respectively, and followed by conductions of monocyte adhesion assay, detection of reactive oxygen species (ROS) generated, and analysis of inflammation-related molecular expression. Our results showed that the monocyte adhesion rate (MDR), which was represented as the level of cell inflammation, was not affected by LSS within 24 h in the absence of nicotine. As compared to the shearing cultures without nicotine, the MDR of the group treated by both nicotine and LSS remarkably increased about 1.8- and 2.6-fold (P < 0.05 for each) while the exposure time was set at 12 and 24 h, respectively. Moreover, the inflammation level of HUVECs treated with both LSS and nicotine for 24 h significantly increased about 1.4 fold (P < 0.05) as compared to the group with nicotine alone. These results were consistent with the data obtained from the ROS measurement that the level of ROS generated from the cells with both LSS and nicotine exhibited 2.55- and 4.22-fold (P < 0.05 for each) higher than the groups with nicotine or LSS alone, respectively. To investigate the mechanism of enhanced endothelial inflammation caused by synergistic effect of LSS and nicotine, expressions of intercellular adhesion molecule-1 (ICAM-1) and vascular adhesion molecule-1 (VCAM-1) were examined. We found that expressions of both ICAM-1 and VCAM-1 increased 1.3 and 1.9-fold while cells were exposed to 10-4 M nicotine for 24 h, whereas LSS enabled to further increased the ICAM-1 but decreased the VCAM-1 expression of nicotine-treated HUVECs after sheared with 12 dynes/cm2 for 24 h, This result suggested that LSS plays different roles in modulating ICAM-1 and VCAM-1 expressions in endothelial cells in the presence of nicotine. In summary, we have demonstrated that the level of endothelial inflammation can be significantly increased by synergistic effect of nicotine and LSS, and exhibited that high level of LSS (≥ 12 dynes/cm2) plays a detrimental role in nicotine-mediated endothelial injury.
關鍵字(中) ★ 尼古丁
★ 層流剪應力
★ 內皮細胞
★ 發炎反應
★ 活性氧自由基
關鍵字(英) ★ Nicotine
★ Laminar shear stress
★ Endothelial cells
★ Inflammation
★ Reactive oxygen species
論文目次 摘要 I
ABSTRSCT III
誌謝 V
目錄 VI
圖目錄 VIII
第一章、 緒論 1
1-1 前言 1
1-2 文獻回顧 3
1-2-1吸菸與心血管疾病之關聯性 3
1-2-2尼古丁對於細胞產生活性氧自由基之相關研究 5
1-2-3層流剪應力對於內皮細胞生理影響之相關研究 6
1-2-4尼古丁與細胞發炎反應之相關性研究 7
1-3 研究目的 9
第二章、 實驗材料與研究方法 11
2-1 實驗材料 11
2-1-1實驗儀器 11
2-1-2實驗藥品與試劑 11
2-1-3平行板流道系統 13
2-2 實驗方法 24
2-2-1細胞於流道內受層流剪力尼古丁刺激之實驗操作 24
2-2-2人類臍靜脈內皮細胞之培養 24
2-2-3人類單核球細胞THP-1培養與染色操作 25
2-2-4血球沾黏測定法 25
2-2-5細胞活性氧自由基測量 26
2-2-6西方墨點法 27
2-2-7抗發炎藥物處理 28
2-2-8統計方法 28
第三章、 實驗結果 29
3-1 人類臍靜脈內皮細胞受層流剪應力尼古丁刺激下血球貼附之實驗結果 29
3-2 層流剪應力尼古丁對人類臍靜脈內皮細胞釋放活性氧自由基量之實驗結果 33
3-3 人類臍靜脈內皮細胞在層流剪應力尼古丁刺激下其貼附因子之蛋白質表現 38
3-4 內皮細胞在尼古丁結合層流剪應力刺激下受藥物的作用 40
第四章、 討論 43
第五章、 後續發展與研究方向 46
參考文獻 47
參考文獻 William B. Kannel, Ralph B. D′Agostino, Albert J. Belanger. “Fibrinogen, cigarette smoking, and risk of cardiovascular disease: Insights from the Framingham Study”, American Heart Journal, Vol 113, pp. 1006-1010, November 1987.
Jieun Lee, John P. Cooke. “The role of nicotine in the pathogenesis of atherosclerosis”, Atherosclerosis, Vol 215, pp. 281-283, April 2011.
Minqi Xu, et al. “The influence of nicotine on granulocytic differentiation – Inhibition of the oxidative burst and bacterial killing and increased matrix metalloproteinase-9 release”, BMC Cell Biology, Vol 9, pp. 1506-1520, April 2008.
Yoav Arnson, Yehuda Shoenfeld, Howard Amital. “Effects of tobacco smoke on immunity, inflammation and autoimmunity”, Journal of Autoimmunity, Vol 34, pp. 258-265, May 2010.
Theodoros G. Papaioannou and Christodoulos Stefanadis. “Vascular Wall Shear Stress: Basic Principles and Methods”, Hellenic J Cardiol, Vol 46, pp. 9-15, November 2005.
Hideyuki Yamawaki, et al. “Fluid shear stress inhibits vascular inflammation by decreasing thioredoxin-interacting protein in endothelial cells”, J Clin Invest, Vol 115, pp. 733-738, March 2005.
Takabe W, E. Warabi and N. Noguchi. “Anti-atherogenic effect of laminar shear stress via Nrf2 activation”, Antioxid Redox Signal, Vol 15, pp. 1415-1426, July 2011.
Malek, A. M., et al. “Hemodynamic shear stress and its role inatherosclerosis”. JAMA: The journal of the American MedicalAssociation , Vol 282, pp.2035-2042,December 1999.
Haltmayer M, et al. “Impact of atherosclerotic risk factors on the anatomical distribution of peripheral arterial disease”, Int. Angiol, Vol 20, pp 200-207, April 2001.
Smith F B, et al. “Smoking, haemostatic factors and the severity of aorto-iliac and femoro-popliteal disease”, Thromb. Haemost, Vol 75, pp. 19-24, November 1996.
William B. Kannel, Ralph B, Albert J. “Fibrinogen, cigarette smoking, and risk of cardiovascular disease: Insights from the Framingham Study”, American Heart Journal, Vol 113, pp. 1006-1010, April 1987.
Hua Cai, David G. Harrison. “Endothelial Dysfunction in Cardiovascular Diseases: The Role of Oxidant Stress”, American Heart Journal, Vol 87, pp. 840-844, September 2000.
H van der Vaart, et al. “Acute effects of cigarette smoke on inflammation and oxidative stress: a review”, Thorax, Vol 59, pp. 713-721, October 2004.
P Kubes, M Suzuki, and D N Granger. “Nitric oxide: an endogenous modulator of leukocyte adhesion”, PNAS, Vol 88, pp. 4651-4655, June 1991.
Joseph Loscalzo. “Nitric Oxide Insufficiency, Platelet Activation, and Arterial Thrombosis”, American Heart Journal, Vol 88, pp. 756-762, April 2001.
Jason P. Eiserich, et al. “Formation of nitric oxide-derived inflammatory oxidants by myeloperoxidase in neutrophils”, Nature, Vol 391, pp. 393-397, January 1998.
B G Harbrecht, et al. “Inhibition of nitric oxide synthesis during endotoxemia promotes intrahepatic thrombosis and an oxygen radical-mediated hepatic injury”, JLB, Vol 52, pp. 390-394, October 1992.
Andrew Churg, et al. “Tumor Necrosis Factor-α Is Central to Acute Cigarette Smoke–induced Inflammation and Connective Tissue Breakdown”, American Journal of Respiratory and Critical Care Medicine, Vol 166, pp. 849-854, September 2002.
Ji-Zhou Dong, Serban C Moldoveanu. “Gas chromatography–mass spectrometry of carbonyl compounds in cigarette mainstream smoke after derivatization with 2,4-dinitrophenylhydrazine”, Journal of Chromatography A, Vol 1027, pp. 25-35, February 2004.
Wilbert S Aronow, Stanleyn Rokaw. “Carboxyhemoglobin Caused by Smoking Nonnicotine Cigarettes Effects in Angina Pectoris”, American Heart Journal, Vol 44, pp. 782-788, July 1971.
A.R. Wakade, T.D. Wakade. “Contribution of nicotinic and muscarinic receptors in the secretion of catecholamines evoked by endogenous and exogenous acetylcholine”, Neuroscience, Vol 10, pp. 973-978, October 1983.
指導教授 李宇翔 審核日期 2015-1-27
推文 facebook   plurk   twitter   funp   google   live   udn   HD   myshare   reddit   netvibes   friend   youpush   delicious   baidu   
網路書籤 Google bookmarks   del.icio.us   hemidemi   myshare   

若有論文相關問題,請聯絡國立中央大學圖書館推廣服務組 TEL:(03)422-7151轉57407,或E-mail聯絡  - 隱私權政策聲明