Division of Periodontology, Department of Oral Biological Science, Niigata University Graduate School of Medical and Dental Sciences, Niigata, Japan.
J Periodontol. 2012 Jul;83(7):917-25. doi: 10.1902/jop.2011.110356. Epub 2011 Nov 28.
Methylation status of the cytokine genes may play a role in the pathogenesis of inflammatory diseases, such as rheumatoid arthritis (RA) and chronic periodontitis (CP). This study was undertaken to evaluate whether the DNA methylation profile of the interleukin-6 (IL-6) gene promoter was unique to individuals with RA and CP.
The study participants consisted of 30 patients with RA, 30 patients with CP, and 30 age-, sex-, and smoking status-balanced healthy controls. Genomic DNA isolated from peripheral blood was modified by sodium bisulfite and analyzed for DNA methylation levels of IL-6 gene with direct sequencing. Levels of IL-6 were determined by an enzyme-linked immunosorbent assay.
The region of IL-6 gene promoter from -1200 to +27 bp was shown to contain 19 CpG motifs. The methylation levels of the CpG motif at -74 bp were significantly lower in patients with RA and CP than those in controls (P = 0.0001). Both levels of serum IL-6 and IL-6 production by mononuclear cells were significantly different between individuals with and without the methylation at -74 bp (P = 0.03). The +19 bp motif exhibited differential levels of the methylation among the groups, which was not associated with serum levels of IL-6. The other 17 CpG motifs exhibited comparable levels of the methylation between the groups.
These results suggest that hypomethylated status of a single CpG in the IL-6 promoter region may lead to increased levels of serum IL-6, implicating a role in the pathogenesis of RA and CP.
细胞因子基因的甲基化状态可能在炎症性疾病(如类风湿关节炎[RA]和慢性牙周炎[CP])的发病机制中发挥作用。本研究旨在评估白细胞介素-6(IL-6)基因启动子的 DNA 甲基化谱是否对 RA 和 CP 患者具有特异性。
研究对象包括 30 例 RA 患者、30 例 CP 患者和 30 例年龄、性别和吸烟状况匹配的健康对照者。从外周血中分离基因组 DNA,用亚硫酸氢钠进行修饰,并直接测序分析 IL-6 基因的 DNA 甲基化水平。通过酶联免疫吸附试验测定 IL-6 水平。
IL-6 基因启动子区域从-1200 到+27 bp 包含 19 个 CpG 基序。RA 和 CP 患者的-74 bp CpG 基序的甲基化水平明显低于对照组(P=0.0001)。有和无-74 bp 位甲基化的个体之间血清 IL-6 水平和单核细胞产生的 IL-6 水平均有显著差异(P=0.03)。+19 bp 基序在各组之间表现出不同的甲基化水平,但与血清 IL-6 水平无关。其他 17 个 CpG 基序在各组之间表现出相似的甲基化水平。
这些结果表明,IL-6 启动子区域单个 CpG 的低甲基化状态可能导致血清 IL-6 水平升高,表明其在 RA 和 CP 的发病机制中发挥作用。