臺灣博碩士論文加值系統

謝格連氏症候群(Sjögren’s syndrome)為一種全身性自體免疫疾病，在淚腺及唾液腺等外分泌腺會發現淋巴球的浸潤。主要症狀會導致口腔唾液分泌不足，及眼睛乾燥現象，在臨床檢驗上病患可測得抗SS-A /Ro(60及52 kDa)以及抗SS-B/La(48 kDa)的表現。只具有乾性角膜炎和口乾症，但未合併其他結締組織病變的病患，稱為原發性謝格連氏症，續發性謝格連氏症則常合併其他結締組織病變一起發生。
本研究的目的在探討細胞激素單核苷酸多型性(SNP)與謝格連氏症是否有相關性存在。所探討之細胞激素單核苷酸多型性(SNP)包括介白質-4 C589T，介白質-6 C174G，介白質-10 A1082G，介白質-10 T819C， 介白質-10 A592C，腫瘤壞死因子-a G238A， 腫瘤壞死因子-a G308A。經過統計分析的結果發現，這些位點在病患與健康人之間的表現並沒有明顯的差異性存在。而細胞激素單核苷酸多型性與自體抗體表現比較結果顯示，介白質-10 A592C與患者表現SS-A/Ro的人數之間有相關性存在( p<0.01)。本研究同時偵測這些細胞激素在血漿中的分泌量並加以統計。在介白質-4部分，健康人平均為10.8±27.4 pg/ml，病患則為3.1 ±15.1 pg/ml，兩者之間有差異性存在( p<0.01)。而介白質-6在血漿中的平均分泌量，健康人為13.3±54.9 pg/ml，病患組則為21.4±61.9 pg/ ml。腫瘤壞死因子-a 的分泌量部分，健康人為25±15.5 pg/ml，病患組則為34.7±61.8 pg/ml，兩者間也沒有顯著性差異存在。介白質-10的表現方面，兩組所得結果皆低於可測值。
因此，依據本研究所得的結果顯示，謝格連氏症與細胞激素單核苷酸多型性(SNP)包括介白質-4 C589T，介白質-6 C174G，介白質-10 A1082G，介白質-10 T819C，介白質-10A592C，腫瘤壞死因子- a G238A，及腫瘤壞死因子- a G308A之間，並沒有明顯的直接相關性存在，且台灣族群在這幾種細胞激素多型性所表現出的基因形，和白種人比較有很大的差異。

Sjögren''s syndrome(SS) is a chronic autoimmune disorder characterized
by dryness of the eyes and mouth,which is typically associated with focal lymphocytic infiltrating of glandular tissues and autoantibody responses against the Ro (SSA,60 and 52 kDa) and La (SSB,48kDa) ribonucleo- proteins.The salivary and lacrimal glands are primarily involved,leading to dry eyes (xerophthalmia with keratoconjunctivitis sicca) and mouth (xerostomia) is called primary Sjögren''s syndrome,and those SS patients (5-20%) encountered with other autoimmune rheumatic disease is called secondary Sjögren''s syndrome.
In this study,we try to derermine the association between cytokine SNP
including interlekin-4,interlekin-6, interlekin-10, tumor necrosis factor- a and clinical manifestations. We have investigated 77 Sjögren''s syndrome patients and 99 healthy females have no autoimmune disease feature,and found that IL-4 C589T, IL-6 C174G, IL-10 T819C, IL-10 A1082G,
IL-10 A592C, TNF- a G238A, TNF- a G308A SNPs are not significant when compared with normal control.However,when autoantibody profiles were examined, IL-10 A592C was significantly associated with anti-SS-A/Ro expression ( p<0.01).We also compared the cytokine plasma level of Sjögren''s syndrome patients and control group.Mean of IL-4 plasma level in normal group is 10.8±27.4 pg/ml,and 3.1±15.1 pg/ml of patients.There was significant difference in plasma secretion level between SS patients and normal control( p<0.01). Mean of IL-6 plasma level in normal group is 13.3±54.9 pg/ml,and 21.4±61.9 pg/ml of patients.The plasma secretion level of TNF- a is 25±15.5 pg/ml in normal group, and 34.7±61.8 pg/ml in SS patients. IL-10 plasma level is nondetective in both groups.
Thus,we suggested that IL-4 C589T, IL-6 C174G, IL-10 A1082G, IL-10
T819C, IL-10 A592C, TNF- a G238A, TNF- a G308A SNPs are not
strong determinants of susceptibility to the development of SS. Futhermore, in Taiwanese idividuals, the genotypes are similar with the distribution of South Chinese idividuals reported previously, and have differences with Caucasian in several cytokine SNPs such as IL-6 C174G, and IL-10 A1082G.

目 錄
壹、中文摘要 1
貳、英文摘要 2
參、綜論
一．歷史回顧 4
二．臨床表徵 6
三．單一核苷酸多型性 (SNP) 8
四．細胞激素與基因多型性 9
五．謝格連氏症與細胞激素 12
六．謝格連氏症與先天基因表現之相關性 12
肆、研究動機 14
伍、材料與方法
一．材料 15
1．檢體來源 15
二．方法
1．問卷調查 15
2．分離血漿 15
3．萃取 DNA 15
4．聚合酶連鎖反應(PCR─ polymerase chain reaction ) 16
5．純化 PCR產物 17
6．DNA 定序 18
7．限制片段長度多型性(RFLP─restriction fragment
length polymorphism ) 18
8．酵素結合免疫吸附分析( ELISA─enzyme- linked 20
immunosorbent assay )
9．資料統計分析 22
陸、結果
一．細胞激素基因多型性實驗 23
二．細胞激素基因多型性與謝格連氏症之相關性 23
三．細胞激素在血漿中之濃度值 24
四．病患自體抗體與細胞激素基因多型性之探討 24
柒、討論 25
捌、參考文獻 28
玖、圖表 33
拾、附圖及附表 41
附圖一、謝格連氏症患者眼部臨床表徵
附圖二、謝格連氏症患者眼部臨床表徵
附圖三、Schirmer’s test
附圖四、謝格連氏症患者口部臨床表徵
附圖五、謝格連氏症患者口部臨床表徵
附表一、問卷 44
附表二、不同細胞激素啟動子基因多型性引子序列
附表三、不同細胞激素啟動子基因多型性聚合酶連鎖反應條件
附表四~十、細胞激素啟動子基因定序之序列結果

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