中文摘要：

目的：检测正常人外周血单个核细胞（PBMC）感染2型登革病毒（DENV2）后肿瘤坏死因子α．（TNF-01）基因启动子区域CpG位点的甲基化水平。方法：采用亚硫酸氢盐测序PCR法检测DNA甲基化水平。结果：TNF-α基因启动子区域为-294bp到＋58bp，覆盖11个散在CpG位点；PCR反应后取PCR产物进行琼脂糖凝胶电泳分析显示，扩增序列大小与理论预测相符合；PBMC感染DENV20h和6h在11个甲基化位点中有2个处于甲基化状态，感染12h有6个甲基化位点甲基化。0h、6h和12h的平均甲基化率分别为10．3％、12．1％和25．5％，且0h和12h及6h和12h的甲基化率差异有统计学意义。结论：PBMC感染DENV2后会引起TNF-α基因启动子区域的甲基化水平增加。

英文摘要：

AIM： To examine DNA methylation at CpG sites in the promoter region of tumor necrosis factor-al- pha （TNF-a） gene in dengue virus type 2 （DENV2） -infected peripheral blood mononuclear cells （PBMC）. METHODS： DNA methylation in the promoter region of TNF-a gene was measured by bisulfite sequencing PCR. RESULTS： The pro- moter region of TNF-a gene was from -294 bp to ＋ 58 bp, including 11 CpG sites. The PCR products identified by aga- rose gel electrophoresis were consistent with the theoretical size. Two sites were methylated at 0 h and 6 h and 6 sites were methylated at 12 h in TNF-a gene promoter region in DENV2-infected PBMC. The average methylation rates were 10. 3%, 12. 1% and 25.5% at 0 h, 6 h and 12 h, respectively. Significant differences between 0 h and 12 h and between 6 h and 12 h were observed. CONCLUSION： The DNA methylation in the promoter region of TNF-a gene is increased in DENV2- infected PBMCs.