从小肠ILCs探讨益气健脾活血法治疗2型糖尿病机制的研究

The small intestine, with its special physiological functions such as immunity and endocrine, is widely involved in the regulation of metabolism. Structural and functional abnormality of small intestine is one of the mechanisms for type 2 diabetes mellitus(T2DM). Intestinal immune imbalance is an important link. Innate lymphoid cells (ILCs) are abundant in small intestinal mucosa, and ILCs dysfunction plays an important role in the pathogenesis of diabetes. In traditional Chinese medicine, the function of the small intestine can be attributed to the function of the spleen. Based on the previous research, ShenQi Compound Recipe of “tonifying qi, invigorating spleen and activating blood” has the following effects: regulating intestinal flora, improving the glucose and lipid metabolism disorder, improving inflammatory status, insulin resistance and obesity. This project intends to adopt ShenQi compound as the carrier to explore the mechanism of the treatment of T2DM with the method of tonifying qi, invigorating spleen and activating blood. Taking small intestinal ILCs as the entry point, the single-cell transcriptome sequencing technology was used to study the effect of “tonifying qi, invigorating spleen and activating blood” method on the subtype and gene expression of small intestinal ILCs in KKAy mice. The purposes of the project is to explore the immune molecular network mechanism of intestinal ILCs involved in the regulation of metabolism, to reveal the internal relationship between the treatment effect of ShenQi compound for T2DM, regulation of ILCs, and molecular signaling pathways and targets, to provide a scientific basis for the clinical application of the method of “tonifying qi, invigorating spleen and activating blood” in the treatment of T2DM.

小肠以其特殊的免疫、内分泌等生理功能，广泛参与机体代谢调控。小肠结构和功能异常是2型糖尿病机制之一，小肠免疫失衡是重要环节。固有淋巴样细胞（ILCs）大量存在于小肠粘膜组织，小肠ILCs功能失衡在糖尿病的发病机制中具有重要作用。中医学认为“脾与小肠相通”，小肠功能可归属于“脾”功能的一部分。课题组前期研究发现，益气健脾活血之“参芪复方”能调节患者肠道菌群，改善糖脂代谢紊乱、炎症、胰岛素抵抗和肥胖等。本项目拟采用参芪复方作为探讨益气健脾活血法治疗T2DM机制研究的载体，以小肠ILCs为切入点，通过单细胞转录组测序技术，研究益气健脾活血法对KKAy小鼠小肠ILCs细胞亚型、基因表达的影响。探索小肠ILCs参与调节机体代谢的免疫分子网络机制，揭示益气健脾活血法治疗糖尿病与调控ILCs及其相关分子信号通路、靶点的之间的内在联系，为益气健脾活血法治疗T2DM的临床应用提供科学依据。

参芪复方已广泛用于治疗T2DM。之前的研究表明，参芪复方可以改善糖脂代谢紊乱，缓解炎症反应等。然而，其潜在机制仍不清楚。本项目采用参芪复方作为探讨益气健脾活血法治疗T2DM机制研究的载体。以小肠ILCs为切入点，取回肠组织制备单细胞悬浮液，通过单细胞转录组测序技术研究益气健脾活血法对KKAy小鼠小肠ILCs细胞亚型、基因表达的影响。结果表明，参芪复方可改善T2DM小鼠的糖脂代谢紊乱、肥胖、胰岛素抵抗和炎症。参芪复方治疗T2DM的可能机制与调节回肠ILCs基因表达有关。进一步分析表明，SQC的治疗机制可能主要与其调控细胞内代谢、有机物代谢、基础代谢、单一有机物代谢、脂代谢、酮酸代谢，有机酸代谢过程相关，涉及多条通路，如免疫炎症反应、代谢、氧化磷酸化通路。分析与糖脂代谢相关的AMPK 信号通路，发现8个基因（Ccnd1、Cd36、Gys1、Hnf4a、Mlycd、Pp2cb、Pp2r1a、Stradb）可能是参芪复方治疗T2DM潜在靶点。本研究在探索中医药防治糖尿病糖脂代谢紊乱的疗效及机制的基础上，明确了糖尿病糖脂代谢紊乱“脾气散精障碍”与回肠ILCs结构功能失衡相关这一重要病理环节，认为中医药防治2型糖尿病糖脂代谢紊乱可通过益气健脾活血法以恢复“脾气散精”功能。