从“肠道菌群-宿主”共代谢角度探讨祛瘀生新法调控溃疡性结肠炎免疫状态的机制

Ulcerative colitis is a chronic nonspecific inflammation. Because its pathogenesis is not clear and there is no ideal treatment method, ulcerative colitis has been one of the hot-spot issues in the digestive disease researches.Our team compared normal people with ulcerative colitis patients of intestinal flora using 16S rRNA sequencing technique,and compared the bacterial differences before and after Traditional Chinese Medicine intervention. Previous research has proved that structure of intestinal flora shows differences between normal and ulcerative colitis patients.The proposal is based on clinical effects of the Traditional Chinese Medicine methods of “qu-yu-sheng-xin，QYSX”（ stasis removing and immunoregulation therapy）on ulcerative colitis.Our Data includes TCM QYSX method that can suppress the gut inflammation and regulate immunity conditions. TCM QYSX method also can improve gut microbiota structure of ulcerative colitis ,increases the abundance of probiotics which produces the short chain fatty acid and decreases the abundance of pathogenic bacteria.This proposal is designed to verify our hypothesis that TCM QYSX method can regulate the immunity condition through metabolite of “gut micobiota-host” co-metabolism to suppress inflammation.To explore this,we will use experiments in vitro and in vivo,combined with 16S rRNA sequencing,technique of metabonomics, magnetic bead cell sorting,flow cytometric method.By demonstrating influence and mechanism of QYSX herbs regulating the immunity,the study will enrich our view of treatment of”stasis removing and immunoregulation therapy”.

溃疡性结肠炎是一种慢性非特异性炎症，因其致病机理尚不明确，且无理想的治疗方法，成为消化疾病研究的热点之一。本项目组前期工作中以16S rRNA测序技术比对溃疡性结肠炎患者和正常人的肠道菌群及患者用药干预前后的肠道菌群变化，前期研究证实：正常人和溃疡性结肠炎患者间的肠道菌群结构存在明显差异，且基于“祛瘀生新法”的中药方剂具有抑制肠道炎症，调整溃疡性结肠炎患者的肠道菌群结构，增加有益菌的丰度及产短链脂肪酸细菌的丰度，并降低致病菌的丰度。本申请围绕“祛瘀生新法”是否通过调整“肠道菌群-宿主”共代谢产物的变化引起免疫状态的改善，以此达到抑制炎症的这一科学假说，采用体内实验、体外实验两种模式来进行验证，综合16S rRNA测序、代谢组学技术、细胞分选技术、流式细胞术等方法。通过深入研究祛瘀生新中药在免疫状态调控中的作用和机制，从而为"祛瘀生新"理论治疗溃疡性结肠炎丰富其科学内涵。

溃疡性结肠炎是一种慢性非特异性肠道炎症，因其致病机理尚不明确，且无理想的治疗方法。溃疡性结肠炎的中医药治疗具有疗效良好，副作用少等特点。以往的中医药治疗溃疡性结肠炎的研究多从中医药调节免疫功能入手，却忽视了肠道菌群和代谢物在疾病发生发展过程的作用。本项目采用体内实验、细胞实验两种模式来进行验证祛瘀生新方的抗肠道炎症的作用。结合肠道菌群测序、代谢组学技术、细胞分选技术、流式细胞术等方法，分析免疫状态、肠道菌群、代谢组三者的相关性，从而为"祛瘀生新"理论治疗溃疡性结肠炎丰富其科学内涵。.研究结果：.（1）祛瘀生新方可以改善DSS诱导的UC小鼠的症状、体征及小鼠肠道的病理损伤。（2）DSS诱导的UC小鼠存在肠道菌群失衡的状态，而祛瘀生新方可以调整UC小鼠的肠道菌群结构，增加有益菌的丰度，降低致病菌的丰度。（3）从细胞层面首次证实祛瘀生新方能促进Treg细胞增殖，并且Treg细胞具有抑制Teff细胞增殖效应。（4）从不同组织证实祛瘀生新方可以调控组织内的炎症因子从侧面反映了其抑制肠道炎症。（5）运用代谢组学检测技术从血液、尿液、粪便不同时空代谢的样品进行探究发现色氨酸代谢通路与肠道炎症的发生密切相关，可能是治疗溃疡性结肠炎的关键代谢通路。.本项目研究开展了用祛瘀生新法治疗溃疡性结肠炎与免疫状态、肠道菌群、代谢组学的相关性研究，发现祛瘀生新方、肠道菌群、代谢组学、免疫状态互为影响，形成一个共同影响着肠道炎症状态的网络模式，该项目为证实祛瘀生新方是系统性网络性治疗溃疡性结肠炎提供了理论和实验依据。