基于外泌体miRNA对M2型巨噬细胞极化的调控探讨湿邪致病病程缠绵的病机本质

湿邪属中医外感六淫之一，致病后易出现缠绵难愈病机，导致病程延长。申请人既往研究发现：湿邪环境下血清miR-155表达明显升高，其靶基因在巨噬细胞极化相关信号通路中显著富集，同时发现M2型巨噬细胞极化有关的TGF-β/HDAC3/TIMAP信号轴表达明显变化。但目前对于湿邪是如何通过miR-155介导M2型巨噬细胞极化，继而影响病程的具体机制仍未明确。基于此我们提出如下科学假说：湿邪可能通过促进血清外泌体将miR-155递送至巨噬细胞，进一步调控TGF-β/HDAC3/TIMAP信号轴，从而抑制M2型巨噬细胞极化，致使邪气缠绵，病程延长。为验证该假说，本项目拟敲除miR-155，验证其可能是湿邪影响机体的重要信号；并进一步证明湿邪环境下递送增多的外泌体miR-155可以通过TGF-β/HDAC3/TIMAP信号轴抑制M2巨噬细胞极化，造成机体炎症持续，从而最终阐明湿邪致病缠绵病机的本质。

Damp-pathogen is one of the six external factors which cause diseases in traditional Chinese medicine theory. Damp-pathogen can cause prolonged disease course. Our previous research found that serum miR-155 expression was significantly increased in the wet environment, and its target genes were significantly enriched in immune and macrophage polarization-related signaling pathways. At the same time, M2 macrophage polarization-related TGF-β/HDAC3/TIMAP signaling pathway-related factors were reduced. However, the relevant mechanism of how damp-pathogen affect the polarization of M2 macrophages through miR-155 is still unclear. Therefore, a scientific hypothesis is proposed: Damp-pathogenl may promote the delivery of miR-155 to macrophages by promoting serum exosomes, and then inhibit the polarization of M2 macrophages through the TGF-β/HDAC3/TIMAP signaling pathway, causing inflammation lasting and the prolonged disease course. In order to clarify this problem, this project uses gene chips, exosomal markers and other technologies to clarify that exosomes miR-155 may be an important molecular pathway for damp-pathogen to affect the body. Combining animal experiments with cell experiments, To clarify that increased delivery of exosomes miR-155 in a wet environment can inhibit M2 macrophage polarization through the TGF-β/HDAC3/TIMAP pathway, causing inflammation lasting and the prolonged disease course.