中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (6): 415-417.

• 论著 • 上一篇    下一篇

5-氮杂胞苷与雌二醇对SLE患者CpG基序甲基化状态与DNMT1表达的影响

朱小华1,梁俊1,杨永生1,徐金华2,项蕾红3   

  1. 1. 复旦大学附属华山医院
    2. 上海市复旦大学附属华山医院皮肤科
    3. 上海华山医院皮肤科
  • 收稿日期:2009-08-31 修回日期:2009-11-22 出版日期:2010-06-15 发布日期:2010-06-08
  • 通讯作者: 朱小华 E-mail:alphabetist@163.com
  • 基金资助:

    上海市科委基金资助课题

Effects of 5-azacytidine and estradiol on the methylation of CpG motifs and expression of DNA methyltransferase 1 in patients with SLE

  • Received:2009-08-31 Revised:2009-11-22 Online:2010-06-15 Published:2010-06-08

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摘要：

目的 探讨5-氮杂胞苷与雌二醇对SLE患者与正常人CpG基序甲基化状态与DNA 甲基转移酶1（DNMT1）表达的影响。方法 分离12例SLE患者与11例正常人外周血淋巴细胞，经植物血凝素刺激1天后，分为未处理组、5-氮杂胞苷组、雌二醇组，继续培养3 d后分别用5-甲基胞嘧啶抗体与流式细胞仪检测CpG基序甲基化水平和实时定量逆转录－聚合酶链反应（RT-PCR）分析DNMT1的mRNA表达水平。结果 与非处理组相比，5-氮杂胞苷组SLE患者（t = 18.60，P < 0.01）与正常人（t = 5.63，P < 0.01）的CpG基序甲基化水平均下降。DNMT1表达水平（正常人t = 2.17，P > 0.05，SLE患者t = 1.56，P > 0.05）的下降无统计学意义。雌二醇组SLE患者与正常人的CpG基序甲基化水平（正常人t = 1.93，P > 0.05；SLE患者t = 1.53，P > 0.05）与DNMT1的mRNA（正常人t = 0.11，P > 0.05；SLE患者t = 0.93，P > 0.05）表达水平的差异均无统计学意义。结论 5-氮杂胞苷可抑制CpG基序甲基化状态，其抑制作用的产生与DNMT1表达水平的下降无明显相关性。雌二醇对CpG基序甲基化状态与DNMT1的表达水平的影响无统计学意义。

关键词: 红斑狼疮，系统性, CpG基序甲基化, DNA甲基转移酶, 5-氮杂胞苷, 雌二醇

Abstract:

Objective To investigate the effects of 5-azacytidine and estradiol on the methylation of CpG motifs and expression of DNA methyltransferase1（DNMT1） mRNA in patients with systemic lupus erythematosus （SLE） and normal controls. Methods Peripheral blood lymphocytes isolated from 12 patients with SLE and 11 normal human controls were stimulated with phytohaemagglutinin for one day followed by additional 3-day culture with or without the presence of 5-azacytidine of 1 μmol/L or estradiol of 30 μg/L respectively. Then, the methylation of CpG motifs was detected by flow cytometry using anti-5-methylcytosine antibody, and DNMT1 mRNA expression by real time reverse transcription-PCR. Results After treatment with 5-azacytidine, a decrease was observed in the methylation of CpG motifs, but not in the expression of DNMT1 mRNA in peripheral lymphocytes from patients with SLE （t = 18.60, P < 0.01; t = 1.56， P > 0.05） and in those from the normal controls （t = 5.63, P < 0.01; t=2.17, P > 0.05） compared with untreated lymphocytes. Nevertheless, there were no significant changes in the methylation of CpG motifs or expression of DNMT1 mRNA in lymphocytes from patients with SLE （t = 1.53，0.93, respectively, both P > 0.05） and normal controls （t = 1.93，0.11，respectively, both P > 0.05） after the treatment with estradiol. Conclusions The methylation of CpG motifs is suppressed efficiently by 5-azacytidine, and the suppression is unlikely to be associated with the decrease of DNMT1 mRNA. Estradiol has no significant impact on the methylation of CpG motifs and expression of DNMT1 mRNA in lymphocytes.

Key words: systemic lupus erythematosus, CpG motifs methylation, DNA Methyltransferase, 5-azacytidine, estradiol