健脾补肾、清肠化湿介导MAPK/ERK促进间充质干细胞迁移分化调节免疫、重建溃疡性结肠炎肠黏膜屏障的研究

ulcerative colitis 's decrease of MSCs is one of the mechanisms that lead to immunologic derangement and difficulty in the rebuilding of intestinal barrier,regulating the immune and rebuilding the intestinal barrier by promoting MSCs' migration and differentiation has already been the new way of treating UC.Previous study shows that bu shen jian piqing chang hua shi fang(sheng huang qi、chao bai zhu、bu gu zhi、yi zhi ren、huang qin、huang lian、di yu、mu xiang、zhi gan cao) could obviously improve UC patients' clinical symptoms, inhibit the inflammatory reaction and promote ulcer healing.However,the mechanism of the study is mainly related to the function of inhibiting matruation of DC and rebuilding the tight junction of intestinal epithelium.The mechanism which shows the function of regulating the immune and rebuilding the intestinal barrier still needs further study to prove.This research , which uses UC animal models characterized by decrease of MSCs,immunologic derangement and destruction of intestinal barrier, and experiment in vitro ,is to find out whether tonifying the kidney and strengthening the spleen, clearing the intestine and resolving dampness could promote MSCs' migration and differentiation,regulate the ratio of Treg/Th17, inhibit inflammatory and immune response,promote differentiation of goblet cells, repair tight junction, rebuilding intestinal barrier and to provide a proof for clinical medicine.

间充质干细胞（MSCs）迁移分化减少是溃疡性结肠炎（UC）免疫紊乱、肠黏膜屏障难以修复的重要机制,介导MAPK/ERK通路促进MSCs的迁移分化调节免疫、重建肠黏膜屏障已成为治疗的新途径。前期研究表明健脾补肾,清肠化湿（生黄芪、炒白术、补骨脂、益智仁、黄芩、黄连、地榆、木香、炙甘草）能显著改善UC患者的症状和黏膜愈合率，其作用机制与抑制DC成熟、NF-κB及炎症因子表达、修复肠上皮紧密连接有关，但与促进MSCs迁移分化的相关性及机制有待进一步阐明。本课题利用具有MSCs减少,免疫调节紊乱,肠屏障破坏的UC动物模型，结合体外实验，研究健脾补肾、清肠化湿通过促进MSCs的迁移分化，进而调节Treg/Th17比例，抑制炎症免疫反应；促进向肠杯状细胞分化，修复紧密连接，重建肠黏膜屏障的作用及机制，为临床用药提供科学依据。

溃疡性结肠炎间充质干细胞（MSCs）迁移分化减少是其免疫紊乱、肠黏膜屏障难以修复的重要机制，本研究利用具有MSCs减少及免疫调节紊乱、肠屏障破坏的溃疡性结肠炎(UC)动物模型，结合体外实验，研究健脾补肾、清肠化湿方是否促进MSCs的迁移分化，抑制炎症免疫反应、修复肠黏膜屏障的作用机制。研究内容包括通过体外实验观察健脾补肾、清肠化湿方对MSCs增殖迁移的影响；通过体内实验观察健脾补肾、清肠化湿方对MSCs移植UC模型大鼠迁移定值情况，对结肠黏膜TGF-β、IL-6、IL-17水平的影响，以及对MSCs 移植后UC模型大鼠肠道屏障的影响。研究结果表明：（1）健脾补肾、清肠化湿方体外对MSCs的生长具有促进增殖作用；MSCs移植UC模型大鼠后，健脾补肾、清肠化湿方可以促进其向肠道迁移定值，并分化为肠干细胞；（2）MSCs移植可以减少促炎细胞因子IL-6、IL-17的分泌水平，提高TGF-β的表达水平；健脾补肾、清肠化湿方干预后，效果优于单纯移植组；（4）MSCs可促进UC模型肠杯状细胞生成，增强大鼠结肠组织中粘液基因Muc2、Muc3的表达，修复粘液屏障，中药干预后的MSCs移植组效果优于单纯MSCs移植组，并且随着时间的延长，修复效果更明显。本研究从炎症免疫和肠道屏障途径观察以MSCs为枢纽，健脾补肾、清肠化湿方影响其增殖迁移，从而降低炎症反应、黏膜修复的作用机制，为临床用药提供客观依据。