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  

Infects of Jiedu-quyu-ziyin Decoction on DNA Methylation of CD4^+T Cell and Glucocorticoid Receptor α Expression in MRL/lpr Mice

作　　者：陈红波[1] 赵丹妮[1] 史佳丽[1] 张利棕[2] 寿旗扬[2] 马红珍[1] 范永升[3] 

机构地区：[1]浙江中医药大学附属第一医院,杭州310006 [2]浙江中医药大学动物实验研究中心 [3]浙江中医药大学风湿病研究所

出　　处：《浙江中医药大学学报》2015年第3期167-171,共5页Journal of Zhejiang Chinese Medical University

基　　金：浙江省自然科学基金(Y2110065);国家自然科学基金(81202673);浙江省卫生厅医药卫生计划(2011KYB059)~~

摘　　要：[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)m RNA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m RNA表达明显上升(P<0.05),Gadd45α基因m RNA表达明显下降(P<0.05),GRα基因m RNA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GRαm RNA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。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.

关 键 词：系统性红斑狼疮 DNA甲基化 糖皮质激素受体 解毒祛瘀滋阴方 

分 类 号：R331[医药卫生—人体生理学]