从组蛋白H3K27me3调控Th17分化偏移探讨补肾活血法治疗强直性脊柱炎分子机制

Inflammation is the initiation stage resulting in the disability of Ankylosing Spondylitis. Th17 is the core effective cell playing an important role in the beginning of AS inflammation. Blocking the differentiation from naive CD4+T cells to Th17 cells becomes a new target of international AS treatment. Supported by the National Natural Science Foundation of China continuously，our research group have many years of work accumulated on clinical and molecular mechanism of tonifying shen and activating blood method on AS. As we found that the differentiation of Th17 was restrained by this method from the previous study, we make the methylation of H3K27 as the core point. In view of this，we hypothesize that this method may inhibiting the activity of histone demethylase JMJD3, and/or the activity of methylase EZH2, affecting the expression of H3K27me3 level within the gene promoter of RORγt、STAT3 and IL-17, so as to relief AS inflammation and restraining the differentiation of Th17. This research discusses the mechanism of tonifying shen and activating blood method with the therapeutic target on epigenetic regulation of Th17 differentiation by H3K27me3, using advanced technologies such as Chip、ChIP-qPCR and so on. At the same time, we use the genetic modification techniques to make JMJD3 overexpression and EZH2 silent in vitro to reverse the direction of Th17 differentiation. After that we could observe the impact of chinese medicine to the histone methylation of target genomic H3K27. In this study we explored the scientific connotation about “kidney essence” and the influence of tonifying shen and activating blood method on cell differentiation regulation from the histone methylation modification.

炎症是强直性脊柱炎（AS）致残始动环节，Th17是触发AS炎症的核心效应细胞。封闭初始CD4+T细胞向Th17分化，成为国际AS治疗学新靶标。依托国家自然科学基金滚动支持，补肾活血法治疗AS临床及分子机制研究有多年积累。在前期发现该法抑制Th17分化基础上，本研究以组蛋白H3K27甲基化修饰为切入点，提出“补肾活血法可能通过抑制去甲基化酶JMJD3活性，和/或促进甲基化酶EZH2活性，影响转录因子RORγt、上游信号分子STAT3及下游IL-17的启动子区域H3K27me3水平，抑制Th17分化偏移，发挥缓解AS炎症作用。”采用ChIP、ChIP-qPCR等技术，探讨补肾活血方干预H3K27me3调控的Th17分化机制；同期体外基因修饰JMJD3/EZH2，诱导Th17分化，观察中药对靶基因组蛋白H3K27甲基化的影响。从组蛋白修饰探索“肾藏精”物质基础及补肾活血法调控细胞分化的科学内涵。

炎症是强直性脊柱炎（AS）致残始动环节，Th17细胞是触发AS炎症的核心效应细胞。封闭初始CD4+T细胞向Th17分化，成为国际AS治疗学新靶标。依托国家自然科学基金滚动支持，补肾活血法治疗AS临床及分子机制研究有多年积累。在前期发现该法抑制Th17分化基础上，本研究以来源于AS患者的外周血单个核细胞为研究对象，以组蛋白H3K27甲基化修饰为切入点，研究AS患者去甲基化酶JMJD3活性，甲基化酶EZH2活性，转录因子RORγt、上游信号分子JAK2/STAT3及下游IL-17的启动子区域H3K27me3水平，同期研究补肾活血法和中药单体干预对其影响作用。研究证实，补肾活血法和中药单体雷公藤甲素可能通过抑制组蛋白去甲基化酶JMJD3活性，和/或促进甲基化酶EZH2活性，影响转录因RORγt、上游信号分子JAK2/STAT3及下游IL-17的基因启动子区域H3K27三甲基化水平，抑制Th17分化偏移，发挥抗AS炎症作用。