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

Abstract

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

References

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Hydroxychloroquine reverses ultraviolet ray-induced genomic DNA hypomethylation in CD4+ T cells from patients with systemic lupus erythematosus

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