冠狀動脈疾病是一種多因子影響的慢性發炎疾病，與脂質代謝異常有關，IL-1ra 為發炎反應引發的抗發炎性細胞激素，可有效降低發炎反應所引起的過度傷害，我們進一步假設 IL-1ra 與發炎性媒介物之間的濃度變化失去平衡對此疾病是有影響。在此篇研究中，我們篩選 70 位健康對照組 ( healthy controls )，95位冠狀動脈疾病 ( CAD ) 的患者共 165 人，分析兩組間冠狀動脈疾病的危險因子、脂質、血清中IL-1ra及其它發炎的細胞激素濃度，IL-1ra 基因多型性用 PCR 去分析，並探討 IL-1ra 濃度變化與基因型的關係對此疾病是否有影響。我們的結果顯示： (1) CAD的危險因子中 BMI、收縮壓、抽煙、高血壓、血糖、HDL-膽固醇及三酸甘油酯在 CAD組有顯著較高，HDL-C 和具抗氧化的 bilirubin 濃度 CAD 組比健康對照組顯著減少 ( p < 0.001)； (2) LDL-C / HDL-C，TC / HDL-C，TG / HDL-C最高級別與最低級別比值分析，有意義增加風險 ( 分別為OR = 2.98，p < 0.01；OR = 5.31，p < 0.001；OR = 8.43，p < 0.001 )； (3) CAD 組抗發炎性細胞激素 IL-1ra 濃度比健康對照組顯著增加 ( p < 0.05 )；CAD組的發炎性檢驗指標 hs-CRP、IL-6、白血球總數、嗜中性球比健康對照組顯著增加 ( p < 0.01 )； (4) 二組間 IL-1ra 與危險因子中的血糖濃度、BMI、TG皆呈正相關，與發炎性檢驗指標 hs-CRP，IL-6，WBC 也呈現正相關；但與HDL- 膽固醇、bilirubin 濃度呈負相關 ( p < 0.01) (5) 以 IL-1ra 依濃度高低級別分析，最高濃度級別比最低濃度級別會增加 2.57 倍罹患 CAD的危險 ( Odds ratio，2.57；95% CI，1.12 to 5.91；p = 0.026 )； (6) hs-CRP 與 IL-6 高濃度會比低濃度增加 CAD 的風險 ( OR = 5.86，p < 0.001；OR = 5.79，p = 0.001 )； (7) hs-CRP、IL-6、IL-1ra 濃度的共同效應的交互作用，可能在 CAD 危險性上扮演重要的角色 ( OR = 10.19，p < 0.001 )； (8) IL-1ra allele 2 基因多型性在 CAD 的表現頻率與 IL-1ra 表現量是無相關的。綜合以上研究結果，IL-1ra 濃度高低與罹患 CAD 的危險性是有相關，除了檢測脂質代謝與發炎標幟物 hs –CRP、IL-6，推測血清 IL-1ra 濃度可提供診斷冠狀動脈疾病的指標之一。

Previous studies show that coronary artery disease (CAD) is a multi-factors and chronic inflammatory disease, and is associated with lipid metabolism. IL-1ra is a naturally occurring anti-inflammatory molecules that block the action of IL-1. However, little is known about the imbalance between IL-1ra and inflammatory mediators in CAD. We attempted to investigate the relationships between inflammatory mediators and serum IL-1ra levels in patients with CAD. In 95 patients with angiographically defined CAD, and 70 healthy controls were studied in a case-control manner. Serum levels of cytokines and the risk factor of CAD were examined. Polymorphisms for IL-1ra gene were detected by PCR, and genotypes and allelic frequencies in both groups were compared. Our major finding include: (1) The risk factors such as elevated BMI, systolic BP, smoking, hypertension, blood glucose, and TG was more frequently found in the CAD group than the control group ( p < 0.001). However, the HDL-C and bilirubin were significantly higher in control group than the CAD group. (2) The relative risk of those in the highest quartile of ratio of LDL-C to HDL-C, TC to HDL-C, and TG to HDL-C were significantly elevated. ( OR = 2.98, p < 0.01; OR = 5.31, p <0.001; OR = 8.43, p < 0.001 respectively) (3) Five different inflammatory markers were significantly elevated including IL-1ra, hs-CRP, IL-6, leukocyte count, and neutrophil percentage between healthy controls and CAD patients. ( p < 0.01) (4) Levels of IL-1ra and other variables such as blood glucose, BMI, TG, IL-6, hs-CRP, and leukocyte count has significantly correlated, and were inversed correlation in bilirubin, and HDL-C in all study subjects. ( p < 0.01) (5) In the multiple logistic regression analysis, adjustment was made for variables. The relative risk of CAD for the highest quartile of IL-1ra, as compared with the lowest quartile, had an Odds ratio 2.57 ( 95% confidence intervals, 1.12 - 5.91, p = 0.026 ) increase in risk for CAD. (6) Similar results were obtained hs-CRP, IL-6 in the highest quartile were increase risk for future CAD. ( OR = 5.86 and 5.79 respectively; p < 0.001) (7) The join effect cytokines of hs-CRP, IL-6, IL-1ra concentrations may play important role in CAD risk. ( OR = 10.19, p < 0.001 ) (8) In addition, IL-1ra allele 2 genotype and allelic frequencies were no significant association with increase in IL-1ra with CAD. In conclusion, we find a significant association of elevated IL-1ra levels in the patients with CAD. Thus, these results support the hypothesis that inflammation, anti-inflammation cytokines and lipoprotein metabolism provide a useful marker for predicting the development of CAD events.

論文審定書
致謝辭
目錄
圖表目次
中文摘要________________________________________i
英文摘要______________________________________iii
英文縮寫表_____________________________________iv
一、序言________________________________________1
冠狀動脈疾病形成________________________________2
IL-1系統________________________________________3
IL-1ra 基因多型性_______________________________5
C反應蛋白 ( C-reactive protein；CRP )___________6
細胞介白素- 6 ( Interleukin-6；IL-6 )___________7
膽紅素 ( Bilirubin )____________________________8
膽固醇 ( Cholesterol )_________________________10
高密度脂蛋白 ( High density lipoprotein；HDL __11
三酸甘油脂 ( Triglycerides；TG )_____________________12
低密度脂蛋白 ( Low density lipoprotein；LDL )__________13
二、實驗材料和方法
檢體收集_______________________________________14
實驗架構_______________________________________15
DNA萃取 ( Genomic DNA extraction )__________________16
PCR ( Polymerase chain reaction )___________________17
電泳 ( Electrophoresis )____________________________19
ELISA ( Enzyme-linked immunosorbent assay )__________21
比色法 ( Colorimetric test )__________________________23
散射比濁法 ( Nephelometry )________________________24
生化自動化分析檢測_______________________________25
統計分析_______________________________________25
三、結果
危險因子在兩組間的差異___________________________26
脂質與脂蛋白於兩組間的表現量______________________26
脂質障礙的評估__________________________________27
發炎與抗發炎標幟物在兩組間的表現___________________27
IL-1ra 與其他變數之間相關性________________________28
IL-1ra濃度高低對罹患冠狀動脈疾病之影響______________29
hs-CRP，IL-6與冠狀動脈疾病之相對危險性_____________30
hs-CRP、IL-6、IL-1ra濃度的共同效應對CAD危險性影響___30
IL-1ra 多型性基因型分布頻率及蛋白質表現______________31
四、討論_______________________________________33
五、參考文獻____________________________________40
六、表_________________________________________46
Table 1. Clinical and biochemical characteristics of coronary artery disease and control subjects
Table 2. Base-line serum levels of lipid markers
Table 3. Risk of coronary artery disease events according to quartile of lipid / HDL-C ratios
Table 4. A list of inflammatory markers in controls and coronary artery disease patients
Table 5. Correlations of other variables with IL-1ra
Table 6. Relative risk of coronary artery disease events according to base-line serum levels of IL-1ra
Table 7. High serum concentrations of IL-1ra are associated with increased coronary artery disease risk
Table 8. Relative risk of coronary artery disease events according to base-line serum levels of inflammation marker
Table 9. The join effect cytokines of hs-CRP, IL-6, IL-1ra concentrations combine in coronary artery disease risk
Table 10. Distribution of Interleukin-1 receptor antagonist genotypes for patients suffering coronary artery disease and also healthy control subjects
Table 11. The effect of IL-1ra genotypes on the IL-1ra levels
七、圖_________________________________________57

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