目的:探讨燃煤污染型地方性氟中毒（简称燃煤型氟中毒）患者外周血DNA甲基化模式及甲基化差异基因，为氟化物诱导机体损伤的发病机制研究提供依据。方法:采用病例-对照研究方法，在贵州省六盘水水城县辖内常年敞灶燃用高氟煤的陡箐乡选择10例重度氟中毒患者为病例组，并在同地区以木柴为主要燃料的花嘎乡选择10例无氟中毒症状者为对照组。采集两组人群外周血，采用简并代表性亚硫酸氢盐测序（RRBS）技术，分析外周血全基因组DNA甲基化模式（n n = 4）、DNA差异甲基化区域（differentially methylated region，DMR），结合DMR差异程度（logn 2Ratio值）和KEGG通路富集分析筛选甲基化差异基因；采用实时荧光定量PCR，验证候选甲基化差异基因的mRNA表达水平（n n = 10）。n 结果:甲基化模式分析结果显示，病例组与对照组基因DNA的所有C碱基甲基化水平分别为（61.53 ± 0.59）%、（62.48 ± 1.53）%；其中，CG位点甲基化水平分别为（63.75 ± 0.65）%、（64.36 ± 1.01）%，CHG位点分别为（13.79 ± 0.72）%、（16.69 ± 4.06）%，CHH位点分别为（25.12 ± 1.72）%、（29.77 ± 3.97）%。与对照组比较，病例组患者分布在不同常染色体上的DMR共有1 000段；其中19号染色体最多，为104段。DMR相关基因978个，其中高甲基化基因265个、低甲基化基因713个；KEGG通路富集分析结果显示，甲基化差异基因主要参与细胞新陈代谢、癌症发生、磷脂酰肌醇3激酶-蛋白激酶B（PI3K-AKT）等信号通路；结合DMR差异程度，以高甲基化的琥珀酸脱氢酶复合物黄素蛋白亚基A假基因3（SDHAP3，logn 2Ratio = 3.487）和低甲基化的核因子κB抑制剂激酶调节亚基γ（IKBKG，logn 2Ratio =-4.436）为候选基因。病例组与对照组人群SDHAP3（0.54 ± 0.08、1.00 ± 0.00）、IKBKG mRNA表达水平（1.32 ± 0.39、1.00 ± 0.00）比较，差异均有统计学意义（n F = 22.94、15.09，n P < 0.01或< 0.05）。n 结论:燃煤型氟中毒患者与对照人群基因组中存在大量的甲基化差异基因，其中髙甲基化的SDHAP3及低甲基化的IKBKG可能参与了燃煤污染型氟化物诱导的机体损伤。“,”Objective:To explore the DNA methylation patterns and methylation differential genes of patients with coal-burning-borne endemic fluorosis, and to provide a basis for study of the pathogenesis of fluoride-induced body injury.Methods:A case-control study was conducted in Shuicheng County, Liupanshui, Guizhou Province, ten patients with severe fluorosis were selected as the fluorosis group in Douqing Township, where people burning high fluorine coal in open range all year round; and ten people without fluorosis phenotype were selected as the control group in Huaga Township, where firewood was the main fuel. Peripheral blood samples were collected from the two groups of people. Reduced representation bisulfite sequencing (RRBS) technique was used to detect the whole genome DNA methylation pattern (n n = 4) and DNA differentially methylated region (DMR), the DMR differential degree (logn 2Ratio) and KEGG pathway enrichment analysis were used to screen the methylation differential genes, and real-time PCR was used to verify the mRNA expression levels of the candidate methylation differential genes(n n = 10).n Results:The methylation pattern analysis results showed that the methylation levels of all C bases in the genome DNA of the fluorosis group and the control group were (61.53 ± 0.59)% and (62.48 ± 1.53)%, respectively; among them, the methylated levels at CG sites were (63.75 ± 0.65)%, (64.36 ± 1.01)%, at CHG sites were (13.79 ± 0.72)%, (16.69 ± 4.06)%, and at CHH sites were (25.12 ± 1.72)%, (29.77 ± 3.97)%. Compared with the control group, patients in the fluorosis group had 1 000 DMR distributed on different autosomes; and the chromosome 19 was the most with 104 segments. There were 978 DMR-related genes, including 265 hypermethylation genes and 713 hypomethylation genes; KEGG pathway enrichment analysis showed that methylation differential genes were mainly involved in cell metabolism, cancers, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) and other signaling pathways; combined with the differential degree of DMR, the hypermethylated succinate dehydrogenase complex flavoprotein subunit A pseudogene 3 (SDHAP3, logn 2Ratio = 3.487) and hypomethylated nuclear factor κB inhibitor kinase regulatory subunit γ (IKBKG, log n 2Ratio =-4.436) were selected as the candidate genes. There were statistically significant differences in the mRNA expression levels of SDHAP3 (0.54 ± 0.08, 1.00 ± 0.00) and IKBKG (1.32 ± 0.39, 1.00 ± 0.00) between fluorosis group and control group (n F = 22.94, 15.09, n P < 0.01 or < 0.05).n Conclusion:There are a large number of methylation differential genes in the genomes of patients with coal-burning-borne endemic fluorosis and controls, the hypermethylated SDHAP3 and hypomethylated IKBKG may be involved in fluoride induced body injury.