解毒祛瘀滋阴方对MRL/lpr小鼠CD4+T细胞DNA甲基化的影响及与GRα表达的相关性研究

摘要

Objective] To investigate the effects of Chinese herbs“Jiedu-quyu-ziyin decoction”on the genomic DNA methylation level and glucocorticoid receptorα (GRα) expression in MRL/Lpr mice. [Methods] Twenty-four MRL/lpr mice(six-week old) were divided into two groups(control group and herb group). Mice spleen CD4+ T cells were separated through immunomagnetic beads. The global genomic DNA methylation levels of CD4+ T cells were measured through commercial kit. The mRNA expression levels of DNMT1, Gadd45α and GRα were detected through real-time quantification PCR. [Results] Eight weeks later, the global DNA methylation level of CD4+T cells in herb group mice was higher than that of control group mice(P<0.05). The mRNA expressions of DNMT1 and GRα in CD4+ T cells of herb group mice were higher than that of control group mice(P<0.05), respectively. Gadd45αmRNA expression in CD4+T cells of herb group mice was lower than that of control group mice(P<0.05). [Conclusion] Jiedu-quyu-ziyin decoction could improve global DNA methylation level of CD4+ T cells in MRL/lpr mice, it also upregulated GRα mRNA expression, but there was no direct correlation between them. We speculated that Jiedu-quyu-ziyin decoction may fulfill its DNA methylation regulation effects through other genes, not GRα.%[目的]探讨解毒祛瘀滋阴方对MRL/lpr小鼠CD4+T细胞基因组DNA甲基化水平的影响及与糖皮质激素受体α（GRα)之间的关系。[方法]将24只6周龄SPF级MRL/lpr狼疮小鼠随机平均分为对照组和中药组。免疫磁珠技术分离出脾脏CD4+T细胞，采用MethyflashTM总体DNA甲基化试剂盒检测CD4+T细胞基因组DNA甲基化水平，实时定量PCR技术检测GRα、DNA甲基化转移酶1（DNMT1）、生长停滞与DNA损伤诱导蛋白α（Gadd45α）mRNA表达水平。[结果]同对照组比较，中药组MRL/Lpr小鼠CD4+T细胞基因组DNA整体甲基化水平明显升高（P<0.05），DNMT1基因mRNA表达明显上升（P<0.05），Gadd45α基因mRNA表达明显下降（P<0.05），GRα基因mRNA表达明显升高（P<0.05）。[结论]解毒祛瘀滋阴方可以升高MRL/lpr狼疮小鼠CD4+T细胞基因组DNA甲基化水平，同时可以上调GRαmRNA表达，但二者之间没有直接联系。解毒祛瘀滋阴方可能是通过其他效应基因实现DNA甲基化调控作用，而不是通过GRα基因。