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研究生:陳英州
研究生(外文):Ying-Chou Chen
論文名稱:唐氏症合併痛風患者之蛋白質體分析
論文名稱(外文):Proteomic Analysis of Down’s Syndrome Patients with Gout.
指導教授:潘台龍
指導教授(外文):Tai-Long Pan
學位類別:碩士
校院名稱:長庚大學
系所名稱:臨床醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2006
畢業學年度:94
語文別:中文
論文頁數:54
中文關鍵詞:唐氏症痛風蛋白質體
外文關鍵詞:Down’s syndromeGoutHPRTproteomics tools
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背景:本研究是以分子生物學與蛋白質體分析法探討唐氏症合併痛風患者HPRT( hypoxanthine-guanine phosphoribosyl transferase) 表現暨血漿蛋白質之改變,進行其機轉以及特性之探索。
方法:採集唐氏症合併痛風患者及一般痛風患者和同齡正常人對照組、及痛風患者之血液檢體以 RT-PCR 及 HPLC-linked 方法來偵測 HPRT 基因 與其酵素活性之表現,並以二維電泳合併質譜分析來合理地解釋有關唐氏症病人血液尿酸量增多之病理機轉與模式研究。
結論:有兩位唐氏症合併痛風性關節患者其 HPRT 表現量略低於正常人, 但仍在正常範圍;另外,由這兩位唐氏症合併痛風患者血清蛋白之圖譜顯示其肝球蛋白( Haptoglobulin )之 α-鏈以及脂蛋白酶元 A-1( Apo-A1) 表現量皆明顯較正常對照組為低,以上結果以西方免疫墨點法進一步確認。
結論:結果顯示與腎臟功能不全有關的肝球蛋白其低表現量可能影響尿酸分泌而導致痛風,而脂蛋白酶元 A-1( Apo-A1) 之低表現量顯示唐氏症患者有脂質代謝缺損且與唐氏症合併痛風病人之痛風性關節炎有正相關。
Background: In this study, the expression of hypoxanthine-guanine phosphoribosyl transferase (HPRT) in Down’s syndrome patients with gout (DS/G) was determined, and the possible underlying mechanisms were characterized with proteomic tools.
Methods: Blood samples from DS/G patients, control and gout patients (without DS), were recruited from the rheumatology outpatient clinic. Baseline enzyme assays were recorded and RT-PCR used to identify the expression of the HPRT gene. 2-D electrophoresis and mass spectrometry were utilized to determine a plausible explanation concerning the pathogenetic mechanisms leading to the increase in uric acid levels in DS patients.
Results: Two DS patients were diagnosed with gouty arthritis. Their HPRT enzyme activity was found to be slightly lower than that of the normal control. The level of HPRT expression was also slightly decreased in DS/G patients compared with the control. Serum protein profiles of these two DS/G patients revealed that haptoglobinαchain and apolipoprotein A1 (ApoA1) were both significantly downregulated. Protein expression was validated by immunoblot.
Conclusion: Our results revealed that their low level of haptoglobin, related to renal dysfunction, may have affected uric acid excretion and led to gout. However, decreased ApoA1 revealed a positive correlation between defective lipid metabolism and gouty arthritis in DS/G patients.
目次
指導教授推薦書 ii
口試委員審定書 iii
授權書 iv
誌謝 v
目錄 vi
圖表目錄 viii
中文摘要 x
英文摘要 xi
第一章 簡介
1.1 研究背景 1
1.2 痛風簡介 2
1.2.1 何謂尿酸及高尿酸血症之定義 2
1.2.2 痛風的定義 4
1.2.3 台灣痛風的現況 5
1.2.4 痛風的症狀 5
1.2.5 HPRT( Hypoxanthine-guaninephosphoribosyltransferase)簡介 7
1.3 唐氏症簡介 7
1.3.1 唐氏症臨床症狀和特徵 8
1.3.2 唐氏症和高尿酸血的關係 10
1.3.3 唐氏症和痛風的關係 11
1.3.4 假說(Hypothesis) 11
1.3.5 策略(Strategy) 12
第二章 材料與方法 13
第三章 實驗結果 18
第四章 討論 20
第五章 未來願景 24
參考文獻 25
圖表 38
附錄
Publications 50
圖表目錄

圖一. Chromosome analysis of Case 1 showed trisomy 21 39
圖二 De novo and salvage pathway synthesis of purine 40
圖三 Urate transporter-1(URAT1) in human kidneys 41
圖四. mRNA expression of HPRT in Down’s syndrome patients with gout 42
圖五. Two-dimensional electrophoresis patterns of plasma proteins from healthy control (N), gout (G) and Down’s syndrome with gout (DS/G) 43
圖六. 圖五之第二蛋白質點之胜肽質譜指紋(PMF)分析及資料庫之蛋白質序列比對後顯示為Apo A1 44
圖七. 圖五之第三蛋白質點之胜肽質譜指紋分析及資料庫之蛋白質序列比對後顯示為Haptoglobulin 45
圖八. The validation of protein levels on haptoglobin alpha chain and ApoA1 through Western blotting 46

表一. Clinical characteristics of the young gout patients 47
表二. The clinical characteristics of uric acid and HPRT activities
between patients and the controls 48
表三. List of identified protein spots 49
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