氧化壓力生物指標和OGG1、ABCA1基因多形性與冠狀動脈心臟病之相關性

Association between oxidative-stress markers and polymorphisms of OGG1 and ABCA1 genes with coronary heart disease

黃如君(Ru-Jiun Huang)

指導教授 ： 葉志清

臺北醫學大學/公共衛生暨營養學院/公共衛生學系暨研究所/碩士(2013年)

摘要

背景介紹：冠狀動脈心臟病為已開發國家主要死因，台灣衛生署2012年死因顯示，心臟疾病位居第二位。冠心病受遺傳與環境因子共同影響，而氧化壓力也扮演重要的角色。氧化壓力對DNA羥化的產物為8-hydroxy-2’-deoxyguanosine (8-OHdG)，而8-isoprostane是脂質過氧化之主要產物。8-oxoguanine glycosylase 1 (OGG1)是一種DNA修復酵素，負責切除DNA上的8-OHdG。ATP-binding cassette-transporter (ABCA1)基因於膽固醇逆運轉途徑中負責重要角色，可調節血漿高密度脂蛋白膽固醇(high-density lipoprotein cholesterol, HDL-C)的濃度。
研究目的：探討8-OHdG和8-isoprostane兩種氧化壓力生物標記與冠心病的相關性，也分析OGG1基因多形性與8-OHdG濃度、ABCA1基因多形性與8-isoprostane濃度對冠心病風險是否有交互作用。
材料方法：本研究以醫院為基礎的橫斷式病例對照研究。研究對象為1999年11月至2004年6月在基隆長庚醫院接受心導管門診檢查之民眾。病例組為具有一條以上冠狀動脈阻塞超過50％者，有445位；對照組為不具有冠狀動脈阻塞者，有204位，合計649位研究對象納入分析。使用Haploview軟體從HapMap資料庫選取OGG1和ABCA1基因之標籤單核苷酸多形性(tagging single nucleotide polymorphisms, Tag SNPs)，基因多形性分析利用Sequenom MassARRAY分析系統，血漿中8-OHdG和8-isoprostane濃度則利用Enzyme-linked immune sorbent assay (ELISA)方法測得。使用邏輯斯迴歸模式分析OGG1和ABCA1基因型及其單體型(haplotype)與冠心病之相關性，並計算其勝算比(odds ratio, OR)與95%信賴區間(confidence interval, CI)。
研究結果：ABCA1 rs2066715帶GA+AA型比起GG型得冠心病的OR為1.62(95% CI=1.05-2.49)，ABCA1 rs2230806帶GA+AA型比起GG型得冠心病的OR為0.54(95% CI=0.34-0.88)。進一步分析ABCA1單體型與冠心病的關係，發現ABCA1單體型為GGTT者相較於GATT者有顯著降低34%的冠心病風險(OR=0.66, 95% CI=0.47-0.92)。而OGG1基因的基因型或單體型與冠心病風險無顯著相關性。另外，8-OHdG和8-isoprostane生物標記或其與OGG1和ABCA1基因間的交互作用並不會影響冠心病的危險性。
結論：ABCA1基因的基因型和單體型與冠心病的風險有關，而OGG1基因與冠心病風險無關。氧化壓力生物標記並未與冠心病有相關性。兩個基因多形性與氧化壓力標記濃度無關。

關鍵字

冠心病、氧化壓力、OGG1基因、ABCA1基因、8-OHdG、8-isoprostane

並列摘要

Objectives:
Coronary heart disease (CHD) is the most prevalent disease in developed countries and the second leading cause of deaths in 2012 in Taiwanese population. Genetic and environmental factors have been related to CHD. Oxidative stress also plays an important role in this disease. The main products resulting from oxidative stress of DNA hydroxylation is 8-hydroxy-2’-deoxyguanosine (8-OHdG) and that of lipid peroxidation is 8-isoprostane. 8-oxoguanine glycosylase 1 (OGG1) is a DNA repair enzyme that excises 8-OHdG from DNA. The ATP-binding cassette-transporter (ABCA1) gene involving in reverse cholesterol transport (RCT) regulates the plasma concentration of high-density lipoprotein cholesterol (HDL-C).
Aim:
The aim of this study was to investigate the association between CHD risk and oxidative-stress biomarkers of 8-OHdG and 8-isoprostane. We also evaluated the interactions between the OGG1 polymorphism and 8-OHdG levels and between the ABCA1 polymorphism and 8-isoprostane levels as well for the risk of CHD.
Methods:
This was a hospital-based cross-sectional case-control study. Participants were recruited at Chang Gung Memorial Hospital in Keelung from January 1999 to June 2004. Cases were angiographically diagnosed with the present of > 50% stenosis in one of the coronary arteries, and controls were with the normal coronary arterises. A total of 649 patients, including 445 cases and 204 controls, were included in our study. The Tag SNPs of the OGG1 and ABCA1 genes in the Han Chinese population were selected using the Haploview software with the International HapMap Project database were genotyped by Sequenom MassARRAY system. The levels of 8-OHdG and 8-isoprostane were determined using the Enzyme-linked immune sorbent assay (ELISA). Logistic regression analysis was used to calculate the odds ratio (OR) and 95% confidence interval (CI) for the OGG1 and ABCA1 genes associated with the risk of CHD.
Results:
The rs2066715 GA+AA genotypes in ABCA1 gene were associated with a significantly increased risk of CHD (OR=1.62, 95%CI=1.05-2.49) compared with the GG genotype. In contrast, the ABCA1 rs2230806 GA+AA genotypes were associated with a significantly decreased risk of CHD (OR=0.54, 95% CI=0.34-0.88), compared with the GG genotype. In haplotype analysis for ABCA1 gene, the risk of CHD was kown for subjects with the GGTT haplotype than those with the GATT haplotype (OR=0.66, 95%CI=0.47-0.92). Neither genotype nor haplotype in OGG1 gene were related to CHD. In addition, the biomarkers of 8-OHdG and 8-isoprostane were not associated with the CHD risk even in interaction with the OGG1 and ABCA1 genes.
Conclusion:
Our study indicates that the risk of CHD is associated with genetype and haplotype in ABCA1 gene, not in OGG1 gene. The oxidative-stress biomarkers have no association with CHD risk.

並列關鍵字

Coronary heart disease ； Oxidative stress ； OGG1 ； ABCA1 ； 8-OHdG ； 8-isoprostane

參考文獻

1. WHO. Cardiovascular diseases (CVDs); 2012.

2. 行政院衛生署. 民國101年死因統計年報; 2013.

3. Boonstra J, Post JA. Molecular events associated with reactive oxygen species and cell cycle progression in mammalian cells. Gene 2004;337:1-13.

4. Maxwell SR. Prospects for the use of antioxidant therapies. Drugs 1995;49(3):345-61.

5. Klungland A, Rosewell I, Hollenbach S, Larsen E, Daly G, Epe B, et al. Accumulation of premutagenic DNA lesions in mice defective in removal of oxidative base damage. Proc Natl Acad Sci U S A 1999;96(23):13300-5.