imDC-Dex参与肠道黏膜免疫耐受拮抗UC的分子机制及健脾益肠散的调节效应

Ulcerative colitis (UC) is a clinical refractory disease, dendritic cells (DC) participated in the UC intestinal mucosal immune and inflammatory responses, and DC secrete body (Dex) is one of the vesicles transport mechanism of mucosal immune. A new study has found that immature DC (imDC) secrete body (imDex) induction of immune tolerance of UC antagonism UC inflammation of the progress. Group preliminary study confirms that the Jian pi yi chang Powder regulating DC and inflammatory cytokines mediated the UC intestinal mucosal immune imbalances and put forward hypothesis, its effect may be mediated via imDC - Dex specificity protein the UC immune tolerance. This project intends to animal in vivo and in vitro cell experiment, combining with electron microscope DC structure and Dex morphology, immunohistochemistry and RT-PCR, Western-Blot method to detect imDC - Dex involved in protein Alix, TSG101, hsc70, LFA - 1, ICAM - 1, CDla, CD83, CD40, CD80, CD86 transcription and expression. In order to make clear imDC - Dex mediated UC intestinal mucosal immune tolerance mechanism, prevention and treatment of UC to Jian pi yi chang Powder provide new pathways and targets, responsible for immune regulation of traditional Chinese medicine treatment of UC accumulating some experience.

溃疡性结肠炎(UC)为临床难治病，树突状细胞(DC)参与了UC肠道黏膜免疫和炎症反应，而DC外泌体(Dex)是黏膜免疫的囊泡运输机制之一。最新研究发现未成熟DC（imDC）分泌的外泌体(imDex)诱导了UC的免疫耐受拮抗UC的炎症进展。课题组前期研究证实健脾益肠散调节了DC与炎症因子介导的UC肠道黏膜免疫失衡，进而提出假说，其作用可能是通过imDC-Dex特异性蛋白介导了UC免疫耐受。本项目拟通过在体动物和离体细胞实验，采用电镜观察DC结构和Dex形态，免疫组化、RT-PCR、Western blot法检测imDC-Dex相关蛋白Alix、TSG101、hsc70、LFA-1、ICAM-1、CDla、CD83、CD40、CD80、CD86的转录与表达，以明确imDC-Dex介导UC肠黏膜免疫耐受机制，为健脾益肠散防治UC提供新的作用途径和靶点，可望为UC的中医药免疫调节治疗积累一些经验。

课题组前期研究证实了健脾益肠散调节DC与炎症因子介导的UC肠道黏膜免疫失衡，由此，我们提出假说，其作用可能是通过imDex特异性蛋白介导了UC免疫耐受。课题组通过整体动物实验和离体细胞实验，研究发现:（1）健脾益肠散降低UC模型大鼠血清中ICAM-1的含量;（2）健脾益肠散降低UC模型大鼠ICAM-1、HSP70、CD40、CD86阳性表达（免疫组化法检测）;（3）健脾益肠散降低UC模型大鼠ICAM-1、HSP70、CD40蛋白表达（Western Blot法检测）;（4）健脾益肠散降低UC模型大鼠ICAM-1、HSP70、CD40 mRNA表达（RT-PCR法检测）;（5）健脾益肠散降低DC细胞CD83蛋白表达（Western Blot法检测）;（6）健脾益肠散降低DC细胞CD83 mRNA表达（RT-PCR法检测）;（7）健脾益肠散降低DC细胞CD40 mRNA表达（RT-PCR法检测）;（8）健脾益肠散降低DC细胞ICAM-1 mRNA表达（RT-PCR法检测）;（9）系统总结了陈慈煦教授诊治脾胃病学术思想的传承和创新;（10）基于“脑-肠-菌轴”理论探讨陈慈煦教授脾胃病用药思想。本项目通过整体动物实验和离体细胞实验，初步明确了imDC-Dex相关蛋白介导UC肠黏膜免疫耐受的分子机制；揭示了健脾益肠散调控imDex及其相关蛋白介导UC肠黏膜免疫耐受机制的作用特点与规律；为健脾益肠散临床治疗溃疡性结肠炎提供了科学依据，初步阐发了健脾益肠散的作用机制，为溃疡性结肠炎的中医药免疫调节治疗方面做了有益的探索。