基于组蛋白H3赖氨酸27修饰的针灸对溃疡性结肠炎IL-9R调节机制研究

It is crucial that aberrant modification of H3 lysine 27 and activation of IL-9 receptor signaling may be related to ulcerative colitis which induced by inflammation of the intestinal mucosal barrier damage. Focus on epigenetic modification and inflammation, we will use tissue morphology, molecular biology, high-throughput sequencing and bioinformatics to study in the experimental animal. For one thing，to confirm that abnormal expressions of the major media IL-9, IL-9R and transcription factor PU.1, IRF4. They may be related to the intestinal mucosal barrier in ulcerative colitis. Acupuncture and moxibustion may inhibit IL-9 receptor signaling, and H3K27 modification is the key factor in ulcerative colitis. For another, to clarify it may be underlying mechanism that role of H3K27 modification on transcriptional regulation and IL-9R gene expression in ulcerative colitis by acupuncture. Acupuncture and modification could protect the intestinal mucosal from inflammation and repair the intestinal mucosal barrier damage through inhibition inflammatory and regulation H3K27 modification, and expect to provide the scientific evidence for effect mechanism of acupuncture therapy for ulcerative colitis.

组蛋白H3赖氨酸27位点（H3K27）异常修饰，及IL-9R信号异常是导致溃疡性结肠炎肠黏膜屏障损伤引发炎症的关键环节之一，本课题在针灸治疗有效的基础上，以表观遗传修饰调控免疫炎症分子为研究切入点，应用组织形态学、分子生物学、高通量测序ChIP-seq及生物信息学等技术与方法，拟从在体动物实验研究：①证实介导溃疡性结肠炎及损伤修复的主要介质IL-9和IL-9R表达异常，其转录因子IRF4和PU.1异常激活，引起肠黏膜屏障损伤是导致结肠发生溃疡的主要环节之一，针灸可抑制IL-9受体信号而调节机体免疫；证明组蛋白H3K27位点修饰是参与溃疡性结肠炎病理的重要调控方式；②初步阐明针灸可能是通过组蛋白H3K27修饰翻译后转录调控IL-9的异常表达调节机体免疫进而保护肠黏膜屏障，控制溃疡性结肠黏膜炎症，以期为针灸治疗溃疡性结肠炎的效应机制提供科学依据。

溃疡性结肠炎（Ulcerative colitis, UC）是炎症性肠病的一种，是临床疑难杂病之一，针灸在控制病情活动、维持缓解结肠炎症和防治并发症方面具有良好疗效。研究证实表观遗传修饰在UC的病情发生、发展中具有重要作用，参与机体的固有免疫和特异性免疫炎症。基于前期研究成果，本项目应用RNA-seq、ChIP-seq、Western blot、Elisa、RT-qPCR、免疫荧光和生物信息学等技术和方法，研究针灸对溃疡性结肠炎大鼠的干预效应，并选择与炎症反应密切相关的分子IL-9和IL-9R，表观遗传学组蛋白修饰H3K27乙酰化和H3K4二甲基化修饰，探讨针灸对UC大鼠结肠炎症和结肠IL-9、IL-9R、PU.1、IRF4、CD4等表达的调节，以及针灸对UC大鼠结肠组织的基因表达谱和组蛋白修饰H3K4me2在免疫因子启动子区域富集作用的影响，阐释针灸对UC大鼠免疫调节的表观遗传修饰机制，为针灸免疫调节机制的阐明提供科学的实验资料。