基于lncRNA-mRNA调控网络的艾灸治疗溃疡性结肠炎的免疫调节机制

Ulcerative colitis (UC) is a refractory disease, moxibustion therapy has a therapeutic advantage. LncRNAs are closely related to the incidence of UC. LncRNA BC012900 overexpression in colonic epithelial cells significantly inhibited cell proliferation, which involves in UC pathogenesis. The aim of this project is to explore lncRNA-mRNA regulatory network, to investigate the role of LncRNA BC012900 and ANRIL transcription factor STAT3/NF-κB and immunoregulation in UC by using the technology and methods of modern molecular biology. To clarify the mechanism of lncRNA BC012900 and ANRIL in intestinal inflammation and the immunomodulatory mechanism of moxibustion in UC by using the siRNA technology. It is suggested that the expression of lncRNAs may be regulated by moxibustion, which acting on transcription factors and inhibiting expression of proinflammatory cytokines from the positive and negative strategies. Moxibustion could relieve intestinal mucosal injury and inflammation and achieve the purpose of prevention and treatment in UC, to provide scientific experimental basis and theoretical foundation for the study of UC inflammatory mechanisms of moxibustion.

溃疡性结肠炎（UC）是一种难治性疾病，艾灸疗法具有疗效优势。晚近，发现lncRNA与UC发病密切相关，lncRNA BC012900在结肠上皮细胞过表达显著抑制了细胞增殖，可能是其参与UC发病的关键环节。本项目拟从UC结肠lncRNA-mRNA调控网络为切入点，应用现代分子生物学等技术与方法，旨从lncRNA BC012900和ANRIL，转录因子STAT3和NF-κB及免疫调节的角度，结合siRNA技术进一步明确lncRNA BC012900和ANRIL在UC肠道炎症中的作用机制，以及艾灸对UC结肠lncRNAs BC012900和ANRIL的免疫调节机制。拟从动物实验方面，利用正反两种策略证实艾灸或是通过影响lncRNAs表达，继而作用转录因子，抑制促炎症因子的转录，有效控制UC肠黏膜损伤及炎症，达到防治UC之目的，以期为艾灸治疗溃疡性结肠炎及临床应用提供实验资料和科学依据。

溃疡性结肠炎（UC）是一种难治性疾病，艾灸疗法具有疗效优势。项目采用RNA-seq高通量测序技术检测lncRNA与mRNA的表达谱，运用生物信息学技术分析和比较艾灸对lncRNA-mRNA互作网络的调控作用，筛选并初步验证了艾灸调控lncRNA表达进而抑制UC大鼠结肠黏膜炎症的关键机制。研究发现并证实：①隔药灸能通过抑制转录因子STAT3与NF-κB p65的磷酸化，降低促炎症因子TNF-α、IFN-γ、IL-1β的蛋白表达，调节Occludin、claudin、ZO-1等结肠黏膜屏障相关蛋白，可能是艾灸保护UC大鼠结肠黏膜屏障功能的免疫调节机制之一。②艾灸能影响UC大鼠结肠组织lncRNA与mRNA表达谱，涉及的主要通路与肠黏膜免疫相关；艾灸能升高结肠组织LOC108352929的lncRNA表达水平，降低Phf11、RT1-N3的mRNA表达水平，可能是艾灸保护UC结肠黏膜屏障机制之一。③体外实验明确了敲低EGCs中LOC108352929的lncRNA表达，TNF-α的mRNA表达显著增加，或是UC结肠黏膜损伤的关键环节;艾灸的抗炎免疫作用，可能是通过lncRNA LOC108352929发挥抑制UC大鼠结肠TNF-α表达作用的。综上所述，揭示了艾灸通过抑制转录因子STAT3与NF-κB p65的磷酸化，降低促炎症因子TNF-α、IFN-γ、IL-1β的蛋白表达，调节Occludin、claudin、ZO-1等结肠黏膜屏障相关蛋白，以及升高结肠组织LOC108352929的lncRNA表达水平，降低Phf11、RT1-N3的mRNA表达水平，升高TNF-α的mRNA表达，缓解UC肠黏膜损伤和炎症，保护肠黏膜屏障功能，为针灸防治UC及临床应用提供实验资料和科学依据。