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研究生: 劉瑾妮
Chin-Ni Liu
論文名稱: 促發炎細胞激素啟動子多型性與台灣帕金森氏症感受性的相關性研究
Studies of association of inflammatory cytokine promoter polymorphisms and expression levels with the susceptibility of Taiwanese Parkinson’s disease
指導教授: 黃基礎
Hwang, Ji-Chuu
李桂楨
Lee, Guey-Jen
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2006
畢業學年度: 94
語文別: 中文
論文頁數: 58
中文關鍵詞: 帕金森氏症介白素-1介白素-6介白素-8促發炎細胞激素
英文關鍵詞: Parkinson's disease, IL-1, IL-6, IL-8, proinflammatory cytokines, SNPs
論文種類: 學術論文
相關次數: 點閱:289下載:13
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  • 帕金森氏症(PD)是一種與年齡相關的神經退化性疾病。目前的相關研究認為帕金森式症成因可能是多種因素共同交互影響產生的結果,包括遺傳因素、環境因素及神經性發炎都可能影響帕金森氏症的發病進程。本研究分析台灣帕金森氏症患族群及正常人族群的IL-1β、IL-6和IL-8基因啟動子區域上的單一核苷酸位點之多型性(single nucleotide polymorphism; SNP)變異,比較正常人與帕金森氏症患者的IL-1β C-511T、IL-6 G-174C和IL-8 T-251A啟動子多型性的差異,以及其不同的基因型與帕金森患者開始發病年齡(age- at-onset)、性別的相關性分析,但並未發現此三種細胞激素與PD發病有相關性。另外選取帶有特定TNF-α -1031基因型(TT或CC)之帕金森患者的淋巴細胞株,以內毒素Lipopolysaccharide (LPS)誘發細胞發炎反應後,藉RT-PCR與同步定量PCR實驗方法,分析TNF-α mRNA表現量,發現各淋巴細胞株對於TNF-α的感受性也有差異。若以cytokine protein array的技術,在促使細胞發炎情況下檢視多種細胞激素的表現量,則發現PD病患之淋巴細胞株經LPS處理後,較正常人之淋巴細胞株釋放出更多的 thrombpoietin、EGF及TNF-β。

    Parkinson's disease (PD) is a neurodegenerative disorder related with aging. The pathogenesis of sporadic PD is thought to be the result of interactions of multiple factors, including inherited factors, environmental factors and neuroinflammation. In the present study, single nucleotide polymorphisms (SNPs) of the promoter region of interleukin-1β gene (IL-1β, C-511T), interleukin-6 gene (IL-6, G-174C) and interleukin-8 gene (IL-8, T-251A) were evaluated for the association with the risk of Taiwanese PD in a case-control study using PCR-restriction enzyme assay. However, the overall genotype and allele distributions at these three sites showed no significant difference between PD cases and controls, even after stratification by age or sex. In addition, RT-PCR and real-time PCR quantitation of LPS-induced TNF-α production in two PD lymphoblastoid cells carrying the TNF-α -1031TT or CC genotype displayed differential expression. The cytokine protein array experiment further revealed that higher level of thrombpoietin, EGF and TNF-β might be induced in response to LPS stimulation in PD lymphoblastoid cells as compared to control lymphoblastoid cells.

    目錄 目錄………………………………………………………………………Ι 中文摘要…………………………………………………………………ΙV 英文摘要…………………………………………………………………V 圖表次……………………………………………………………………VI 壹、緒論 一、帕金森氏症……………………………………………………1 二、病因學…………………………………………………………2 (一)環境因素………………………………………………………2 (二)遺傳因素………………………………………………………5 三、中樞神經發炎與帕金森氏症的相關性…………………………6 四、促發炎細胞激素啟動子基因多型性與帕金森氏症 (一)IL-1β………………………………………………………………………………………………………8 (二)IL-6……………………………………………………………10 (三)IL-8……………………………………………………………10 五、研究動機及目的…………………………………………………11 貳、研究材料與方法 一、研究樣品…………………………………………………………13 二、基因組DNA的萃取………………………………………………13 三、聚合酵素連鎖反應(PCR) ……………………………………14 四、基因型分析(genotyping)……………………………………14 五、內生性IL-1β、IL-8 mRNA表現量檢測………………………15 六、淋巴細胞株TNF-α mRNA表現量檢測 (一)淋巴細胞株培養……………………………………16 (二)淋巴細胞處理………………………………………17 (三)核糖核酸(RNA)的萃取與定量……………………17 (四)反轉錄聚合酵素連鎖反應…………………………17 (五)同步定量PCR (Real-time PCR)………………18 七、淋巴細胞株cytokine antibody array (一)淋巴細胞處理………………………………………19 (二)Cytokine表現量檢測………………………………20 參、結果 一、IL-1β基因………………………………………………………21 (一)IL-1β 啟動子多型性分析...…………………………21 (二)PD患者內生性IL-1β mRNA表現量……………………22 二、IL-8基因……………………………………………………………22 (一)IL-8啟動子多型性分析...……………………………23 (二)PD患者內生性IL-8 mRNA表現量………………………24 三、IL-6基因多型性分析………………………………………………24 四、淋巴細胞株經LPS誘導後TNF-α mRNA的表現情形………………25 五、淋巴細胞株Cytokine Antibody Array表現量檢測…………26 肆、討論 一、IL-1β啟動子多型性分析……………………………………………28 二、IL-6和IL-8啟動子多型性分析……………………………………29 三、刺激物誘導TNF-α的表現會因細胞株、劑量及處理時間而有所差 異………………………………………………………………………29 四、Cytokine antibody array篩檢其他可能影響PD致病的cytokine標 的…………………………………………………………………………30 伍、文獻參考………………………………………………………………33 附錄..........................41

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