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研究生:黃瀅云
研究生(外文):Huang, Ying-Yun
論文名稱:S100A8蛋白、醣化終產物接受器(RAGE)與基質金屬蛋白酶-1(MMP-1)之基因多形性與膝部退化性關節炎之相關性
論文名稱(外文):Association of S100A8, Receptor for Advanced Glycation End Products and Matrix Metalloproteinase-1 Gene Polymorphisms with Knee Osteoarthritis
指導教授:蘇遂龍蘇遂龍引用關係
指導教授(外文):Su, Sui-Lung
口試委員:蘇遂龍周雨青李建和
口試委員(外文):Su, Sui-LungChou, Yu-ChingLee, Chien-Ho
口試日期:2011-05-23
學位類別:碩士
校院名稱:國防醫學院
系所名稱:公共衛生學研究所
學門:醫藥衛生學門
學類:公共衛生學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:126
中文關鍵詞:S100A8蛋白基質金屬蛋白酶-1醣化終產物接受器退化性關節炎基因多形性
外文關鍵詞:MMP-1S100A8RAGEpolymorphismOsteoarthritis
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退化性關節炎為已開發國家中最常見的關節疾病,過去文獻指出軟骨降解及發炎反應於退化性關節炎扮演重要角色。S100蛋白及醣化終產物接受器(RAGE)與發炎反應的調控有關,並會影響基質金屬蛋白酶(MMPs)的產生,而基質金屬蛋白酶-1(MMP-1)會造成軟骨之降解,可能影響退化性關節炎的發展。此外,過去研究顯示醣化終產物接受器、S100A8蛋白及基質金屬蛋白酶-1的基因多形性(SNP)會影響配體接合或轉錄調控,故本研究目的為探討此三者之基因多形性與退化性關節炎之相關性。
本研究採用病例對照研究法,選取三軍總醫院就診骨科接受關節置換術的病患共207位作為病例組;而三軍總醫院健檢之民眾經醫師以X光片診斷為K/L< 2者中,依病例組分佈情形來進行年齡、性別之配對,選取配對好之207位作為對照組。以聚合酶連鎖反應(PCR)及限制酶片段長度多形性(RFLP)的方法進行基因型判定,再以SPSS統計軟體進行分析。
研究結果顯示,BMI於病例組與對照組間達統計顯著水準,其危險對比值為1.38( 95% CI: 1.27-1.49)。而基因多形性的結果中,在調整年齡、性別及BMI後,RAGE -429 T/C和557G/A之基因多形性,於病例組與對照之分布達統計上顯著水準。於RAGE -429 T/C,以TT型為參考組,CT型之危險對比值為0.42( 95%CI: 0.21-0.85),而將CT型、TT型合併成一組與TT型做比較,其危險對比值為0.48( 95%CI: 0.25-0.93)。而於RAGE 557G/A,以GG型為參考組,AA型之危險對比值為2.78( 95%CI: 1.02-7.63),並將A對偶基因與G對偶基因做比較,其危險對比值為1.49( 95% CI: 1.04-2.13)。而其他部分則顯示與膝部退化性關節炎無相關性存在。
結論,在台灣人中BMI為膝部退化性關節炎重要之危險因子,且RAGE的基因多形性可能影響膝部退化性關節炎,可視為膝部退化性關節炎的易感性基因。

Osteoarthritis (OA) is the most common arthritic disease in developed countries. The inflammation of cartilage has known to play an important role in OA. The S100 and receptor for advanced glycation end products (RAGE) has been demonstrated to participate in regulation of inflammation, even in production of matrix metalloproteinases (MMPs). MMP-1 has known to degrade cartilage which may result in development of OA. Moreover, polymorphisms of RAGE, S100A8 and MMP-1 were shown to have a marked effect on ligand binding and transcription regulating. We investigated the association of gene polymorphisms with OA.
We performed a case control study. 207 patients were diagnosed with knee osteoarthritis and underwent total knee replacement. The control group involved 207 individuals who had standard x-rays of the knee joints to confirm K/L < 2 and matched by age and sex. All subjects were drawn from Tri-Service General Hospital. Polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay in all subjects. Data was analyzed using SPSS statistical software.
In this study, there was significant difference in BMI between the case and control groups. After adjusted for age, sex and BMI of the subjects, RAGE -429 T/C and 557G/A showed significant differences between case and control groups. The frequencies of CT and CT+CC genotypes of RAGE -429 T/C were significantly lower in OA patients than in the control group ( OR=0.42, 95%CI: 0.21-0.85; OR=0.48, 95%CI: 0.25-0.93, respectively). In addition, The frequencies of AA genotype and A allele of RAGE 557G/A were significantly higher in OA patients than in the control group (OR=2.78, 95%CI: 1.02-7.63; OR=1.49,95%CI: 1.04-2.13, respectively). However, there was no association of S100A8 and MMP-1 polymorphisms with susceptibility to OA.
Our data suggested that BMI is a risk factor of OA and RAGE polymorphisms is one of the likely candidate determinants of OA.

目錄
第一章 前言 1
第一節 研究背景及重要性 1
第二節 研究目的 3
第二章 文獻探討 4
第一節 退化性關節炎 4
第二節 退化性關節炎有關之調控機制 10
第三節 醣化終產物接受器(RAGE) 13
第四節 S100蛋白 19
第五節 基質金屬蛋白酶(MMP) 24
第三章 材料與方法 31
第一節 研究設計與對象 31
第二節 研究資料之收集 31
第三節 研究架構 35
第四節 實驗方法 36
第四章 統計分析 48
第五章 結果 50
第一節 研究對象之人口背景資料 50
第二節 基因多形性哈-溫平衡 51
第三節 醣化終產物接受器(RAGE)基因多形性與退化性關節炎之相關性 53
第四節 S100A8基因多形性與退化性關節炎之相關性 62
第五節 基質金屬蛋白酶-1(MMP-1)基因多形性與退化性關節炎之相關性 65
第六節 RAGE連鎖不平衡檢定與單套型分析 74
第七節 MMP-1連鎖不平衡檢定與單套型分析 76
第八節 RFLP分析基因多形性 78
第六章 討論 85
第一節 探討退化性關節炎相關之人口學變項 85
第二節 探討RAGE基因多形性 86
第三節 探討S100A8 基因多形性 93
第四節 探討MMP-1基因多形性 95
第五節 研究限制 103
第七章 結論與未來展望 104
第八章 參考文獻 106
附錄一 127


圖目錄
圖一、關節軟骨中造成軟骨細胞死亡及退化性關節炎產生與老化有關的細胞及細胞外基質的改變 12
圖二、RAGE整體構造 14
圖三、RAGE基因之基因多形性分布情形 17
圖四、S100蛋白於細胞內、外調控反應 20
圖五、多種基質金屬蛋白酶之結構 26
圖六、膝部X光攝影之結果所判定的Kellgren/ Lawrence (K/L)等級 34
圖七、研究架構 35
圖八、RAGE基因多形性之連鎖不平衡檢定 75
圖九、MMP-1基因多形性之連鎖不平衡檢定 77
圖十、RAGE -429 T/C 基因多形性之瓊脂凝膠電泳圖 78
圖十一、RAGE -374 T/A 基因多形性之瓊脂凝膠電泳圖 79
圖十二、RAGE 557 G/A 基因多形性之瓊脂凝膠電泳 80
圖十三、S100A8 rs3795391 A/G基因多形性之瓊脂凝膠電泳圖 81
圖十四、MMP-1 -1607 1G/2G基因多形性之瓊脂凝膠電泳圖 82
圖十五、MMP-1 -755 G/T基因多形性之瓊脂凝膠電泳圖 83
圖十六、MMP-1 -519 A/G基因多形性之瓊脂凝膠電泳圖 84

表目錄
表一、多配體接受器RAGE與其相關疾病 15
表二、RAGE之基因多形性與病理狀態之相關性 18
表三、MMP-1 -1607之基因多形性與疾病之相關性 30
表四、Kellgren/ Lawrence (K/L)分級 33
表五、PCR引子序列 41
表六、PCR試劑配置(一) 42
表七、PCR試劑配置(二) 43
表八、PCR反應之條件 44
表九、RFLP 反應試劑與條件 46
表十、退化性關節炎邏輯式迴歸單變項分析 50
表十一、對照組基因多形性之哈-溫平衡檢定 51
表十二、退化性關節炎病例組與對照組RAGE -429 T/C基因型、對偶基因頻率、顯隱性模式分布情形 55
表十三、退化性關節炎病例組與對照組RAGE -374 T/A基因型、對偶基因頻率、顯隱性模式分布情形 58
表十四、退化性關節炎病例組與對照組RAGE 557 G/A 基因型、對偶基因頻率、顯隱性模式分布情形 61
表十五、退化性關節炎病例組與對照組S100A8 rs3795391 A/G基因型、對偶基因頻率、顯隱性模式分布情形 64
表十六、退化性關節炎病例組與對照組MMP-1 -1607 1G/2G基因型、對偶基因頻率、顯隱性模式分布情形 67
表十七、退化性關節炎病例組與對照組MMP-1 -755 G/T基因型、對偶基因頻率、顯隱性模式分布情形 70
表十八、退化性關節炎病例組與對照組MMP-1 -519 A/G基因型、對偶基因頻率、顯隱性模式分布情形 73
表十九、RAGE單套型與退化性關節炎的相關性分析 75
表二十、MMP-1單套型與退化性關節炎的相關性分析 77
表二十一、基因多形性在不同種族中對偶基因之分布頻率 102

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