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目的近期研究发现FOXP3基因CNS2区的去甲基化水平可反映调节性T细胞水平。调节性T细胞在动脉粥样硬化中起保护作用。本研究旨在应用焦磷酸测序的方法检测该基因片段的甲基化水平,评估冠心病患者调节性T细胞水平的变化。方法和结果本研究纳入114位急性冠脉综合征患者以及11位冠脉造影正常者。提取外周血DNA,应用焦磷酸测序的方法检测的FOXP3基因CNS2区域甲基化水平。试验发现,冠心病患者FOXP3基因甲基化分析所得到的调节性T细胞水平均较对照者显著降低(正常对照组11.97%±2.01%,急性冠脉综合征组9.79%±1.77%;P<0.001);分别根据冠脉病变血管的严重程度和Gensini评分的三分位数将冠心病患者分组分析,结果提示各组冠心病患者较正常对照者的调节性T细胞均显著减少。FOXP3-CNS2的去甲基化水平与冠心病的严重程度呈反比(r=-0.206,P<0.05)。在去除传统危险因素的影响后,两者相关性降低,且无显著性差异。结论本研究显示冠心病患者的去甲基化FOXP3所代表的调节性T细胞水平显著降低,调节性T细胞的减少和动脉粥样硬化的严重程度尚需进一步证实。
Purpose Recent studies found that the demethylation level of FOXP3 gene CNS2 region can reflect the level of regulatory T cells. Regulatory T cells play a protective role in atherosclerosis. This study aimed to detect the methylation level of this gene fragment by pyrosequencing and evaluate the change of regulatory T cells in patients with coronary heart disease. Methods and Results This study enrolled 114 patients with acute coronary syndrome and 11 patients with normal coronary angiography. Peripheral blood DNA was extracted and methylation level of FOXP3 gene CNS2 region was detected by pyrosequencing. The test found that patients with coronary heart disease FOXP3 methylation analysis of regulatory T cells were significantly lower than the control group (normal control group 11.97% ± 2.01%, acute coronary syndrome group 9.79% ± 1.77%; P < 0.001). According to the severity of coronary artery lesion and the tertile of Gensini, coronary heart disease patients were divided into groups and analyzed. The results showed that all the patients with coronary heart disease had significantly fewer regulatory T cells than the normal controls. The demethylation level of FOXP3-CNS2 was inversely correlated with the severity of coronary heart disease (r = -0.206, P <0.05). After removing the influence of traditional risk factors, the correlation between the two was reduced, with no significant difference. Conclusions This study shows that the level of regulatory T cells represented by demethylated FOXP3 is significantly decreased in patients with coronary heart disease. The decrease of regulatory T cells and the severity of atherosclerosis need to be further confirmed.