乙酰胆碱对巨噬细胞功能调控异常致先天性巨结肠相关性小肠结肠炎的机制研究

先天性巨结肠相关性小肠结肠炎（HAEC）是先天性巨结肠最常见的并发症且是该病患儿死亡的主要原因，但发病机制不明。项目组的前期研究提示HAEC可能与乙酰胆碱（ACh）对未成熟期巨噬细胞（MΦ）活化的抑制作用下降有关。本项目拟采用分子生物学、免疫学、激光显微切割、活体成像、转录组学及生物信息学等研究手段，研究人类和模型鼠HAEC病变肠道乙酰胆碱含量、MΦ分布及组织病理学改变；阐明不同时期M1型MΦ的ACh受体及信号通路特点；比较ACh对不同时期M1型MΦ活化调控作用的异同；探讨不同剂量ACh对不同时期M1型MΦ功能的影响；厘清M1型MΦ活化异常导致的免疫反应紊乱及其对肠上皮细胞损伤。通过以上研究将明确ACh对M1型MΦ活化调控的信号通路及其该通路功能失常在HAEC发病中的作用，为治疗HAEC寻找到新的方向。

Hirschsprung’s disease-associated enterocolitis (HAEC) is the most common cause of death in patients with Hirschsprung’s disease (HD). However, the underlying mechanism of HAEC remains unclear. In our previous studies, we have confirmed that HAEC may be closely associated with the decreased inhibition of activating immature macrophages (MΦ) by acetylcholine (ACh). In the current project, we aim to investigate ACh concentration, distribution of macrophages and pathological changes in the bowels of HD patients and murine model of HD by using methods of molecular biology, immunology, laser capture microdissection，in-vivo imaging, transcription and bioinformatics. Additionally, we propose to explore the characteristics of ACh receptors and their signaling pathways in various stages during activation of M1 MΦ. Furthermore, the function of M1 MΦ in various stages will be observed after being dealt with different doses of ACh. At last, we plan to clarify the pathogenesis of immune response disorder caused by abnormal M1 MΦ over-activation and its effect in damaging intestinal epithelial cells. Our study will be helpful in illustrating the roles of ACh in M1 MΦ activation signaling pathways and in manifesting the dysfunction of the pathways in the pathogenesis of HAEC, which may shed light on novel therapies for HAEC.