miRNA-185 和DNA 甲基转移酶1 在妊娠合并 系统性红斑狼疮患者T 细胞中的表达及其意义

Expression and significance of miRNA-185 in T cells of pregnant patients with systemic lupus erythematosus

摘要:

目的 探讨miRNA-185 和DNA 甲基转移酶1（DNMT1）在妊娠合并系统性红斑狼疮（SLE） 患者T 细胞中的表达及在发病中的作用。方法 采用实时荧光定量聚合酶链反应和蛋白质印迹检测DNMT1 mRNA 和蛋白的表达，核转染后72 h 检测甲基化敏感性基因的水平。并与健康人群比较，分析其表达差异 及意义。结果 结果显示，miRNA-185 在妊娠合并SLE 患者的CD4+T 细胞中上调（P <0.05），其过表达与 DNMT1 mRNA 水平呈负相关。在健康人CD4+T 细胞中，miRNA-185 的过表达会导致DNA 低甲基化、 CD11a 和CD70 基因编码的上调。抑制SLE 患者的CD4+T 细胞中miRNA-185 的表达，会起到DNA 甲基化 逆转作用。结论 miRNA-185 能靶向作用于DNMT1，并导致系统性红斑狼疮患者妊娠期间CD4+T 细胞的 DNA 低甲基化。miRNA-185 可能是SLE 干预治疗的潜在治疗靶点。

关键词:系统性红斑狼疮;妊娠;CD4+T 细胞;miRNA-185 ;DNA 甲基转移酶1 ;表达

Abstract:

Objective To investigate expression of miRNA-185 and DNA methyltransferase 1 (DNMT1) in T cells of pregnant patients with systemic lupus erythematosus (SLE) and potential clinical significance. Methods Expression of miRNA-185 and DNMT1 in T cells was measured by qRT-PCR and Western blot. Methylation level of T cells in SLE patients was determined. Results Expression of miRNA-185 was upregulated significantly in CD4+T cells from patients, which was negatively correlated with DNA methyltransferase 1 (DNMT1) mRNA levels. Overexpression of miRNA-185 in CD4+T cells from healthy donors led to the DNA hypomethylation and up-regulation of CD11a and CD70 genes. Inhibition of miRNA-185 expression in CD4+T cells from patients with SLE caused reverse effects. Target prediction analysis and dual luciferase reporter assays confirmed that DNMT1 was a direct target of miRNA-185. Conclusions This study indicates that miRNA-185 contributes to DNA hypomethylation in CD4+T cells of pregnant patients with SLE by targeting DNMT1,which may represent a potential therapeutic target for SLE.

Key words:systemic lupus erythematosus; pregnancy; CD4+T; miRNA-185; DNA methyltransferase 1; expression