To investigate the effect of bromodomain and extra-terminal (BET) protein inhibitor JQ1 on expression of autoimmune-related genes in CD4+T cells from patients with systemic lupus erythematosus (SLE). Methods: Peripheral CD4+T cells were isolated by positive selection with CD4 microbeads. The percentage of CD4+T cells were detected by flow cytometry. CD4+T cells were treated by JQ1 at 100 nm/L for 6, 24, 48 h. The expression of T cell-related genes was measured by quantitative real-time PCR (qPCR). The secretion levels of cytokines in culture supernatant were measured by ELISA at 48 h. Results: The percentage of CD4+T cells isolated by CD4 microbeads is 97.2%. Compared with the control group, the mRNA expression levels of IFNG, IL-17F, IL-21, CXCR5 and FOXP3 were down-regulated at 6, 24 and 48 h (P<0.05), and IL-17A mRNA level was decreased at 6 and 24 h (P<0.01); while IL-4 mRNA level was up-regulated at 24, 48 h (P<0.01), and TGF-β1 mRNA level was up-regulated at 6 and 48 h (P<0.05) in SLE CD4+T cells treated with JQ1. The secretion levels of IFN-γ and IL-21 in JQ1-treated group were decreased significantly (P<0.05), while the secretion levels of IL-4 and TGF-β were up-regulated compared with control group (P<0.05). Conclusion: JQ1 can reverse the immune dysregulation and improve the immunity homeostasis in CD4+T cells from patients with SLE.
目的：探讨溴结构域(bromodomain and extra-terminal，BET)蛋白抑制剂JQ1对系统性红斑狼疮(systemic lupus erythematosus，SLE)外周血CD4+T细胞中自身免疫相关基因表达的作用。方法：利用磁珠分选获得SLE患者CD4+T细胞，采用流式细胞检测CD4+T细胞纯度。用JQ1(100 nm/L)处理CD4+T细胞6，24，48 h后收集细胞。采用实时荧光定量PCR(quantitative real-time PCR，qPCR)检测不同辅助性T细胞亚类特异性基因的mRNA表达。采用ELISA法检测细胞培养上清中细胞因子的蛋白水平。结果：磁珠分选所得CD4+T细胞百分比为97.2%。与对照组相比，JQ1处理组IFNG，IL-17F，IL-21，CXCR5和FOXP3 mRNA表达水平在6，24，48 h均显著降低(P<0.05)，IL-17A mRNA表达在6，24 h显著下降(P<0.01)，IL-4 mRNA表达在24，48 h显著升高(P<0.01)，TGF-β1 mRNA表达在6，48 h显著升高(P<0.05)。JQ1处理48 h细胞培养上清中IFN-γ，IL-21和IL-17蛋白水平明显下降(P<0.05)，IL-4和TGF-β蛋白水平明显升高(P<0.05)。结论：BET蛋白抑制剂JQ1可纠正SLE患者CD4+T细胞中的免疫调节失衡，促进免疫稳态恢复。.