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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
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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

    Adams JD, Chang ML, Klaidman L (2001) Parkinson's disease-redox mechanisms. Currt Med Chem 8, 809-814.
    Ascherio A, Zhang SM, Hernan MA, Kawachi I, Colditz GA, Speizer FE, Willett WC (2001) Prospective study of caffeine consumption and risk of Parkinson's disease in men and women. Ann Neurol 50, 56-63.
    Bagli M, Papassotiropoulos A, Knapp M, Jessen F, Luise Rao M, Maier W, Heun R (2000) Association between an interleukin-6 promoter and 3' flanking region haplotype and reduced Alzheimer's disease risk in a German population. Neurosci Lett 283, 109-112.
    Barbeau A, Roy M, Cloutier T (1986) Smoking, cancer, and Parkinson's disease. Ann Neurol 20, 105-106.
    Barbosa ER, Limongi JCP, Cummings JL (1997) Parkinson's disease. Psych Clin North Am 20, 769-790.
    Bertram L, Tanzi, RE (2005) The genetic epidemiology of neurodegenerative disease. J Clin Invest 115, 1449-1457.
    Beutler B, Cerami A (1989) The biology of cachectin/TNF-a primary mediator of the host response. Annu Rev Immunol 7, 625-655.
    Beutler B, van Huffel C (1994) Unraveling function in the TNF ligand and receptor families. Science 264, 667-668.
    Blum-Degen D, Muller T, Kuhn W, Gerlach M, Przuntek H (1995) Interleukin-1beta and interleukin-6 are elevated in the cerebrospinal fluid of Alzheimer's and de novo Parkinson's disease patients. Neurosci Lett 202, 17-20.
    Bonifati V, Rizzu P, van Baren MJ, Schaap O, Breedveld GJ, Krieger E, Dekker MC, Squitieri F, Ibanez P, Joosse M (2003) Mutations in the DJ-1 gene associated with autosomal recessive early-onset parkinsonism. Science 299, 256-259.
    Cardenas H, Bolin LM (2003) Compromised reactive microgliosis in MPTP-lesioned IL-6 KO mice. Brain Res 19, 89-97.
    Casper D, Mytilineou C, Blum M (1994) EGF enhances the survival of dopamine neurons in rat embryonic mesencephalon primary cell culture. J Neurosci Res 30, 372-381.
    Castano A, Herrera AJ, Cano J, Machado A (1998) Lipopolysaccharide intranigral Injection induces inflammatory reaction and damage in nigrostriatal dopaminergic system. J Neurochem 70, 1584-1592.
    Crossgrove J, Zheng W (2004) Manganese toxicity upon overexposure. NMR Biomed 17, 544-553.
    D'Amato RJ, Lipman ZP, Snyder SH (1986) Selectivity of the parkinsonian neurotoxin MPTP: toxic metabolite MPP+ binds to neuromelanin. Science 231, 987-989.
    Daurer W, Przedborski S (2003) Parkinson's Disease: Mechanisms and models. Neuron 39, 889-909.
    El-Omar EM, Carrington M, Chow WH, McColl KE, Bream JH, Young HA, Herrera J, Lissowska J, Yuan CC, Rothman N, Lanyon G, Martin M, Fraumeni JF,Rabkin CS (2000) Interleukin-1 polymorphisms associated with increased risk of gastric cancer. Nature 404, 398-402.
    Friedman WJ (2001) Cytokines regulate expression of the type 1 interleukin-1 receptor in rat hippocampal neurons and glia. Exp Neurol 168, 23-31.
    Gayle DA, Ling Z, Tong C, Landers T, Lipton JW, and Carvey PM (2002) Lipopolysaccharide (LPS)-induced dopamine cell loss in culture: roles of tumor necrosis factor-alpha, interleukin-1beta, and nitric oxide. Brain Res Dev Brain Res 133, 27-35.
    Gao HM, Hong JS, Zhang W, Liu B (2003) Synergistic dopaminergic neurotoxicity of the pesticide rotenone and inflammogen lipopolysaccharide:relevance to the etiology of parkinson's disease. J Neurosci 23, 1228-1236.
    Gibb WR, Lees AJ (1991) Anatomy, pigmentation, ventral and dorsal subpopulations of the substantia nigra, and differential cell death in Parkinson's disease. J Neurol Neurosurg Psychiatry 54, 388-396.
    Griffin WS, Sheng JG, Royston MC, Gentleman SM, McKenzie JE, Graham DI, Roberts GW, Mrak RE (1998) Glial-neuronal interactions in Alzheimer's disease: the potential role of a ‘cytokine cycle’ in disease progression. Brain Pathol 8, 65-72.
    Halliwell B, Chirico S (1993) Lipid peroxidation: its mechanism, measurement, and significance. Am J Clin Nutr 57, 715-725.
    Hanke M, Farkas LM, Jakob M, Ries R, Pohl J, Sullivan AM (2004) Heparin-binding epidermal growth factor-like growth factor: a component in chromaffin granules which promotes the survival of nigrostriatal dopaminergic neurones in vitro and in vivo. Neuroscience 124, 757-766.
    Hernan MA, Takkouche B, Caamano-lsoma F, Gestal-Otero JJ (2002) A meta-analysis of coffee drinking, cigarette smoking, and the risk of Parkinson's disease. Ann Neurol 52, 276-284.
    Ho A, Blum M (1997) Regulation of astroglial-derived dopaminergic neurotrophic factors by interleukin-1b in the striatum of young and middle-aged mice. Exp Neurol 148, 348-359.
    Hunot S, Dugas N, Fauchenx B, Hartmann A, Tardien M, Debre P, Agid Y, Dugas B, Hirsch EC (1999) FecR II/CD23 is expressed in parkinson's disease and induced, in vitro, production on nitric oxide and tumor necrosis factor-α in glial cells. J Neurosci 19, 3440-3451.
    Jenner P (2003) Oxidative stress in Parkinson's disease. Ann Neurol 53, s26-s38.
    Kim WG, Mohney RP, Wilson B, Jeohn GH, Liu B, Hong JS (2000) Regional difference in susceptibility to lipopolysaccharide-induced neurotoxicity in the rat brain: role of microglia. J Neurosci 20, 6309-6316.
    Kitada T, Asakawa S, Hattori N, Matsumine H, Yamamura Y, Minoshima S, Yokochi M, Mizuno Y, Shimizu N (1998) Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. Nature 392, 605-608.
    Krizanac-Bengez L, Kapural M, Parkinson F, Cucullo L, Hossain M, Mayberg MR, Janigro D (2003) Effects of transient loss of shear stress on blood-brain barrier endothelium: role of nitric oxide and IL-6. Brain Res 977, 239-246.
    Kruger S, Brandt E, Klinger M, Kreft B (2000) Interleukin-8 secretion of cortical tubular epithelial cells is directed to the basolateral environment and is not enhanced by apical exposure to Escherichia coli. Infect Immun 68, 328-334.
    Lai JC, Minski MJ, Chan, AW, Leung TK, Lim L (1999) Manganese mineral interactions in brain. Neurotoxicology 20, 433-444.
    Langston JW (1987) MPTP: insights into the etiology of Parkinson's disease. Eur Neurol 26 Suppl 1, 2-10.
    Liu J, Zhao ML, Brosnan CF, Lee SC (2001) Expression of type II nitric oxide synthase in primary human astrocytes and microglia: role of IL-1beta and IL-1 receptor antagonist. J Immunol 157, 3569-3576.
    Mattila KM, Rinne JO, Lehtimaki T, Roytta M, Ahonen JP, Hurme M (2002) Association of an interleukin-1beta gene polymorphism (-511) with Parkinson's disease in Finnish patients. J Med Genet 39, 400-402.
    McGeer PL, Yasojima K, McGeer EG (2002) Association of interleukin-1 beta polymorphisms with idiopathic Parkinson's disease. Neurosci Lett 21, 67-69.
    Mehta DS, Wurster AL, Whitters MJ, Young DA, Collins M, Grusby MJ (2003) IL-21 Induces the Apoptosis of Resting and Activated Primary B Cells. J Immuno 170, 4111-4118.
    Miller RJ, Meucci O (1999) AIDS and the brain: is there a chemokine connection? TINS 22, 471-479.
    Mizuno Y, Hattori N, Matsumine H (1998) Neurochemical and neurogenetic correlates of Parkinson's disease. J Neurochem 73, 893-902.
    Moynagh PN, Williams DC, O’neill LAJ (1994) Activation of NF-κB and induction of vascular cell adhesion molecule-1 and intracellular adhesion molecule-1 expression in human glial cells by IL-1. J Immunol 153, 2682-2689.
    Montgomery EJ (1995) Heavy metals and the etiology of Parkinson's disease and othermovement disorders. Toxicology 97, 3-9.
    Morens DM, Grandinetti A, Reed D, White LR, Rosss GW (1995) Cigarette smoking and protection from Parkinson's disease: false association or etiologic clue? Neurology 45, 1041-1051.
    Nishimura M, Mizuta I, Yamasaki S, Ohta M, Kuno S (2000) Influence of interleukin-1beta gene polymorphisms on age-at-onset of sporadic Parkinson's disease. Neurosci Lett 284, 73-76.
    Owen AM (2004) Cognitive dysfunction in Parkinson's disease: the role of frontostriatal circuitry. Neuroscientist 10, 525-537.
    Papassotiropoulos A, Bagli M, Jessen F, Bayer TA, Maier W, Rao ML, Heun R (1999) A genetic variation of the inflammatory cytokine interleukin-6 delays the initial onset and reduces the risk for sporadic Alzheimer's disease. Ann Neurol 45, 666-668.
    Polymeropoulos MH, Lavedan C, Leroy E, Ide SE, Dehejia A, Dutra A, Pike B, Root H, Rubenstein J, Boyer R, et al. (1997) Mutation in the alpha-synuclein gene identified in families with Parkinson's disease. Science 276, 2045-2047.
    Rothwell N, Allan S, Toulmond S (1997) Perspective series: cytokines and the brain. The role of interleukin 1 in acute neurodegeneration and stroke: pathophysiological and therapeutic implications. J Clin Invest 100, 2648-2652.
    Schipper HM (1998) Glial iron sequestration and neurodegeneration. In: HM, S (Eds), Astrocytes in brain ageing and neurodegeneration RG Landes Co, Austin.
    Schipper HM (2004) Brain iron deposition and the free radical-mitochondrial theory of ageing. Ageing Res Rev 3, 265-301.
    Schulte T, Schols L, Muller T, Woitalla D, Berger K, Kruger R (2002) Polymorphisms in the interleukin-1 alpha and beta genes and the risk for Parkinson's disease. Neurosci Lett 21, 70-72.
    Shahi GS, Moochhala SM (1991) Smoking and Parkinson's disease-a new perspective. Rev Environ Health 9, 123-136.
    Skipper L, Farrer M (2002) Parkinson's genetics: molecular insights for the new millennium. Neurotoxicology 23, 503-514.
    Snyder SH, D'Amato RJ (1986) MPTP: a neurotoxin relevant to the pathophysiology of Parkinson's disease. Neurology 36, 250-258.
    Tanner CM, Ottman R, Goldman SM, Ellenberg J, Chan P, Mayeux R, Langston JW (1999) Parkinson disease in twins: an etiologic study. JAMA 281, 341-346.
    Tsao N, Hsu HP, Wu CM, Liu CC, Lei HY (2001) Tumor necrosis factor-alpha causes an increase in blood-brain barrier permeability during sepsis. J Microbio 50, 812-821.
    Vassalli P (1992) The pathophysiology of tumor necrosis factors. Annu Rev Immunol 10, 411-452.
    Valente EM, Abou-Sleiman PM, Caputo V, Muqit MM, Harvey K, Gispert S, Ali Z, Del Turco D, Bentivoglio AR, Healy DG, et al. (2004) Hereditary early-onset Parkinson's disease caused by mutations in PINK1. Science 304, 1158-1160.
    Wang J, Bankiewicz KS, Plunkett RJ, Oldfeld EH (1994) Intrastriatal implantation of interleukin-1: reduction of parkinsonism in rats by enhancing neuronal sprouting from residual dopaminergic neurons in the ventral tegmental area of the midbrain. J Neurosurg 80, 484-490.
    Zimprich A, Biskup S, Leitner P, Lichtner P, Farrer M, Lincoln S, Kachergus J, Hulihan M, Uitti RJ, Calne DB, et al. (2004) Mutations in LRRK2 cause autosomal-dominant parkinsonism with pleomorphic pathology. Neuron 44, 601-607.

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