Histone Modification In Peripheral Blood Cells From Diabetic Patients

AimsTo investigate the global histone modification patterns and the status of histone H3 acetylation at Tumor necrosis factor-a (TNF-a) and cyclooxygenase-2 (COX-2) gene promoter region in peripheral blood mononuclear cells (PBMCs) collected from Type 2 diabetic patients.MethodsGlobal histone H3/H4 acetylation and H3K4/H3K9 methylation in PBMCs from 12 Type 2 diabetic patients and 12 healthy control subjects were quantified by the EpiQuikTM global histone H3/H4 acetylation and H3K4/H3K9 methylation assay kit. The differential expression of the histone modification enzymes and TNF-a mRNA, COX-2 mRNA were measured by real-time polymerase chain reaction (Real-time PCR). Chromatin immunoprecipitation analysis was used to detect the status of histone H3 acetylation at TNF-a and COX-2 gene promoter region.ResultsIncreased global H3K4 methylation (P<0.05), H3 acetylation (P <0.01) and H4 acetylation(P<0.05) was observed in PBMCs from type 2 diabetic patients compared with controls. mRNA level of the histone deacetylases HDAC2, SIRT1 in patients PBMCs was 3.55±0.30 (P <0.01),0.40±0.25 (P<0.05) fold that in healthy controls. mRNA level of the histone acetyltransferases P300, CREBBP in patients PBMCs was 3.11±0.38 (P<0.05),2.78±0.45 (P<0.05)fold that in healthy controls. mRNA level of the histone demethylase KDM3A, KDM5B, LSD1 in patients PBMCs was 0.43±0.43 (P<0.05),3.55±0.30 (P<0.01),0.36±0.35 (P<0.05) fold that in healthy controls. mRNA level of the histone methytransferases SET1 in patients PBMCs was 3.06±0.51 (P<0.05) fold that in healthy controls. TNF-a and COX-2 mRNA was overexpressed in PBMCs from Type 2 diabetic patients compared with normal controls. H3 acetylation at the TNF-a and COX-2 gene promoter region was elevated in PBMCs from Type 2 diabetic patients compared with normal controls.ConclusionAn aberrant expression pattern of histone modification and histone modification enzymes existed in PBMCs of T2DM. H3 acetylation at the TNF-a and COX-2 gene promoter region was elevated. AimsTo investigate global histone modification patterns and the status of histone H3 acetylation at Forkhead box protein 3 (Foxp3) gene promoter regions in CD4+ T cells collected from latent autoimmune diabetes in adults (LADA).MethodsGlobal histone H3/H4 acetylation and H3K4/H3K9 methylation in CD4+ T cells from 16 LADA patients and 16 healthy control subjects were analyzed by the EpiQuikTM global histone H3/H4 acetylation and H3K4/H3K9 methylation assay kits. The differential expression of the histone modification enzymes and Foxp3 mRNA were measured by Real-time PCR. Chromatin immunoprecipitation analysis was used to detect the changes of histone H3 acetylation at the promoter region of Foxp3 gene.ResultsReduced H3/H4 acetylation was observed in the LADA CD4+ T cells compared with the controls (P<0.05). Compared with healthy controls, mRNA levels of the histone deacetylases HDAC2, HDAC5 and SIRT1 in the CD4+T cells of patients were upregulated, mRNA levels of the histone acetyltransferases P300, CREBBP, PCAF in patients CD4+ T cells were downregulated. mRNA level of the histone demethylase LSD1 in patients CD4+ T cells was 0.17±0.13 (P<0.01) fold that in healthy controls. mRNA level of the histone methytransferases SUV39H2 in patients CD4+ T cells was 0.36±0.11 (P<0.05) fold that in healthy controls. Foxp3 mRNA was downregulated in CD4+ T cells from LADA patients (P<0.05). H3 acetylation at the Foxp3 gene promoter region (-501/-416) (-204/-114) was lower in CD4+ T cells from LADA patients compared with normal controls (P<0.01)ConclusionAn aberrant expression pattern of histone modification and histone modification enzymes existed in CD4+ T cells of LADA. H3 acetylation at the Foxp3 gene promoter region (-501/-416) (-204/-114) was decreased. Objective To investigate whether the histone deacetylase inhibitor, trichostatin A (TSA) interfere with H3 acetylation and Foxp3 expression in CD4+ T cells from LADA patients.Methods Peripheral blood mononuclear cells (PBMC) isolated from blood of healthy volunters were prepared using the Ficoll-Hypaque method. CD4+ T cells were isolated using magnetic beads. CD4+ T cells were treated with different concentrations of TSA. Cells were collected after appropriate time, H3 acetylation and the expression of Foxp3 was analyzed by Western-blot.Results Compared with control group, TSA intervention increased H3 acetylation and Foxp3 expression in CD4+ T cells from LADA patients time-dependantly and dose-dependantly.Conclusion H3 acetylation and Foxp3 expression in LADA CD4+ T cells was elaveted by TSA treatment.