羟氯喹逆转紫外线诱导的SLE患者CD4+T细胞DNA低甲基化的研究

摘要/Abstract

摘要：

目的探讨羟氯喹对紫外线诱导的SLE患者CD4+ T细胞基因组DNA低甲基化的作用。方法选择SLE患者组30例，正常人对照组10例。磁珠分选SLE患者组和正常人对照组外周血CD4+ T细胞，311 nm窄谱UVB照射，加入羟氯喹共培养，检测各组间基因组DNA甲基化表达水平。结果 SLE患者组CD4+ T细胞DNA甲基化水平（3.922 ± 2.215）%低于正常人对照组［（10.210 ± 5.573）%，t = 3.450，P = 0.026］；SLE活动组患者CD4+ T细胞经45 mJ/cm2和100 mJ/cm2 UVB照射后DNA甲基化水平为（1.784 ± 1.033）%和（1.932 ± 1.844）%，均显著低于活动期患者未照射组［（3.922 ± 2.215）%，t = 3.000、4.118，P值均 < 0.05］。经100 mJ/cm2 UVB照射后，活动期患者DNA甲基化水平（1.932 ± 1.844）%显著低于稳定期患者照射组［（7.235 ± 3.846）%，t = 2.648，P < 0.05］和正常人对照组［（5.472 ± 5.573）%，t = 3.000，P < 0.05］。SLE活动组T细胞经45和100 mJ/cm2 UVB照射后，加用羟氯喹结果DNA甲基化水平为（4.698 ± 1.948）%和（8.698 ± 3.151）%，均比照射未加羟氯喹组显著升高（t = 4.827、3.184，P值均 < 0.05）；经45 mJ/cm2 UVB照射前后均加羟氯喹组DNA甲基化水平（5.404 ± 2.308）%比照射未加羟氯喹组（1.784 ± 1.033）%显著升高，t = 4.827，P < 0.01。结论羟氯喹可以逆转紫外线诱导的SLE患者CD4+ T细胞DNA低甲基化，羟氯喹对活动期SLE患者更为明显。

关键词: DNA甲基化

Abstract:

Objective To explore the effect of hydroxychloroquine on ultraviolet ray-induced genomic DNA hypomethylation in CD4+ T cells from patients with systemic lupus erythematosus （SLE）. Methods Thirty patients with SLE and 10 normal human controls were enrolled in the study. CD4+ T cells were isolated from these subjects by using magnetic beads, and cultured. Hydroxychloroquine of 50 mg/L was added to the culture medium of CD4+ T cells before or after the exposure to narrow band ultraviolet B （NB-UVB） 311 nm. After additional culture, the levels of genomic DNA methylation in CD4+ T cells were determined with the Imprint Methylated DNA Quantification kit. Results The levels of DNA methylation was lower in SLE patients than in the normal controls ［（3.922 ± 2.215）% vs. （10.210 ± 5.573）%, t = 3.450, P = 0.026］. After exposure to UVB at 45 and 100 mJ/cm2, the DNA methylation level in patients with active SLE decreased from （7.235 ± 3.846）% to （1.784 ± 1.033）% and （1.932 ± 1.844）% respectively （t = 3.000, 4.118, both P < 0.05）. Decreased DNA methylation level was observed in CD4+ T cells from patients with active SLE compared with those from patients with stable SLE and normal human controls ［（1.932 ± 1.844）% vs. （7.235 ± 3.846）% and （5.472 ± 5.573）%, t = 2.648, 3.000, both P < 0.05］ after irradiation with UVB of 100 mJ/cm2. A significant increase in the methylation level was observed in active SLE patient-derived CD4+ T cells treated with hydroxychloroquine following the irradiation with UVB of 45 （4.698% ± 1.948%） and 100 mJ/cm2（8.698% ± 3.151%） compared with those only treated with UVB irradiation （t = 4.827, 3.184, both P < 0.05）, as well as in those treated with hydroxychloroquine before and after the irradiation with UVB of 45 mJ/cm2 compared with those receiving irradiation alone ［（5.404 ± 2.308）% vs. （1.784 ± 1.033）%, t = 4.827, P < 0.01］. Conclusions Hydroxychloroquine can reverse the UVB-induced genomic DNA hypomethylation in CD4+ T cells from patients with SLE, especially in those from patients with active SLE.

Key words: DNA methylation

陈学琴汪国生张敏赵小娟张宏李向培. 羟氯喹逆转紫外线诱导的SLE患者CD4+T细胞DNA低甲基化的研究[J]. 中华皮肤科杂志, 2011, 44(6): 419-422.

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