Zhang Min, Fang Xuan, Wang Guo-Sheng, Ma Yan, Jin Li, Li Xiao-Mei, Li Xiang-Pei
Department of Rheumatology and Immunology, Anhui Provincial Hospital, Anhui Medical University, No. 17 Lujiang Road, Hefei, 230001, People's Republic of China.
Inflammopharmacology. 2017 Apr;25(2):203-210. doi: 10.1007/s10787-017-0321-8. Epub 2017 Feb 11.
In the present study, DNA methylation level of CD4+ T cells exposed to ultraviolet B (UVB) was investigated and its potential mechanisms were also explored.
CD4+ T cells from 12 cases of healthy subjects and 33 cases of SLE patients were isolated and exposed to different dosages (0, 50, 100 mJ/cm) of UVB. Further, SLE patients were divided into two groups: active SLE group (22 cases, SLEDAI scores >4) and inactive SLE group (11 cases, SLEDAI scores ≤4). DNA methylation was evaluated by the Methylamp™ Global DNA Methylation Quantification Ultra Kit. The mRNA and protein expression levels of DNA methyltransferases (DNMT1 and DNMT3A) were detected by real-time PCR and western blot, respectively.
The levels of DNA methylation and DNMT3A mRNA in SLE patients were significantly decreased compared with those in healthy subjects at baseline. After different dosages of ultraviolet irradiation (0, 50 and 100 mJ/cm), DNA methylation levels of CD4+ T cells were all reduced in a dose-dependent manner in three subgroups. Additionally, 100 mJ/cm ultraviolet irradiation in active SLE group contributed to a significant decrease of both DNA methylation and DNMT3A mRNA levels in CD4+ T cells. UVB exposure had no significant effects on expression levels of DNMT1 mRNA and protein and DNMT3A protein.
UVB decreases DNA methylation level of CD4+ T cells in SLE patients probably via inhibiting DNMT3A mRNA expression level, which needs to be further explored.
本研究调查了暴露于紫外线B(UVB)的CD4+ T细胞的DNA甲基化水平,并探讨了其潜在机制。
分离12例健康受试者和33例SLE患者的CD4+ T细胞,使其暴露于不同剂量(0、50、100 mJ/cm)的UVB。此外,将SLE患者分为两组:活动期SLE组(22例,SLEDAI评分>4)和非活动期SLE组(11例,SLEDAI评分≤4)。采用Methylamp™全球DNA甲基化定量超试剂盒评估DNA甲基化。分别通过实时PCR和蛋白质印迹法检测DNA甲基转移酶(DNMT1和DNMT3A)的mRNA和蛋白质表达水平。
与健康受试者基线水平相比,SLE患者的DNA甲基化水平和DNMT3A mRNA水平显著降低。在不同剂量紫外线照射(0、50和100 mJ/cm)后,三个亚组中CD4+ T细胞的DNA甲基化水平均呈剂量依赖性降低。此外,活动期SLE组100 mJ/cm的紫外线照射导致CD4+ T细胞中DNA甲基化和DNMT3A mRNA水平均显著降低。UVB暴露对DNMT1 mRNA和蛋白质以及DNMT3A蛋白质的表达水平无显著影响。
UVB可能通过抑制DNMT3A mRNA表达水平降低SLE患者CD4+ T细胞的DNA甲基化水平,这一机制有待进一步探索。