Tumor necrosis factor-alpha gene promoter methylation in Japanese adults with chronic periodontitis and rheumatoid arthritis

Background and ObjectiveOver-expression of tumor necrosis factor-alpha (TNF-) plays a pathological role in chronic periodontitis (CP) and rheumatoid arthritis (RA), which might be regulated by the epigenetic mechanism. The aim of the present study was to evaluate whether there is a unique methylation profile of the TNF- gene promoter in blood cells of individuals with CP and RA. Material and MethodsThe study participants consisted of 30 Japanese adults with RA (RA group), 30 race-matched adults with CP only (CP group) and 30 race-matched healthy controls (H group). Genomic DNA isolated from peripheral blood was modified by sodium bisulfite and analyzed, by direct sequencing, to investigate DNA methylation of the TNF- gene promoter region. The level of TNF- produced in mononuclear cells stimulated with Porphyromonas gingivalis lipopolysaccharide was determined using ELISA. ResultsTwelve cytosine-guanine dinucleotide (CpG) motifs were identified in the TNF- promoter fragment from -343 to +57bp. The CP group showed a significantly higher methylation rate and frequency at -72bp than the H group (p<0.01). The RA group exhibited significantly higher methylation rates at seven CpG motifs (-302, -163, -119, -72, -49, -38 and +10bp), and significantly higher methylation frequencies at six CpG motifs (-163, -119, -72, -49, -38 and +10bp), than the H group (p<0.01 for all comparisons). The levels of TNF- produced were significantly different between individuals with and without methylation at -163bp (p=0.03). ConclusionThese results suggest that the hypermethylated status of CpG motifs in the TNF- gene promoter in blood cells may be unique to Japanese adults with CP and RA.