小分子化合物作为拮抗剂的筛选及其在肠道炎症性疾病中的应用

摘要：近年来，肠道炎症性疾病的发生率持续上升。肠道炎症性疾病的发病原因及机制尚未明确。有研究表明，肠道黏膜免疫系统异常反应所导致的炎症反应在结肠炎发病中起到重要作用。维甲酸相关孤核受体家族中的RORγt作为T辅助17细胞及3型天然淋巴细胞所共有的转录因子，在T辅助17细胞及3型天然淋巴细胞的分化及相关细胞因子的分泌中起到重要作用，从而影响肠道炎症性疾病的发生。一类小分子化合物可作为维甲酸相关孤核受体家族中的RORγt的拮抗剂影响相关细胞的分化。本文通过T辅助17细胞体外分化，筛选出对T辅助17细胞有效的拮抗剂GSK805及SR1001，将两者应用于固有层淋巴细胞的刺激实验；探究发现，GSK805及SR1001对于小鼠大肠及小肠LPL中3型天然淋巴细胞分泌IL-22均没有影响。若经过进一步的动物实验，可探究拮抗剂体内作用的复杂机制，从而对自身免疫缺陷相关肠道炎症性疾病的靶向治疗奠定基础。48717

毕业论文关键词：拮抗剂；维甲酸相关孤核受体；T辅助17细胞；3型天然淋巴细胞；肠炎

Some kinds of small molecular compounds RORγt antagonist and It's effects and aplications

Abstract:In the recent years, the incidence of inflammatory bowel disease (IBD) continues to rise. At this point, the causes of IBD remains unknown, and more research needs to be down. Studies have shown that inflammatory response caused by the abnormal reactions of the intestinal mucosal immune system plays an important part in the incidence of UC. RORγt works as a transcription factor, and it is shared by Th17 and ILC3. It plays an important role in the differentiation of Th17and ILC3, and the secretion of cytokines. A kind of small molecular compounds can be used as RORγt antagonist and effects the differentiation of related cells. In this paper, GSK805 and SR1001 are selected as effective antagonist through Th17 differentiation in vitro. Then using these two antagonist in stimulating experiment about LPL. We find that neither GSK805 nor SR1001 can inhibit the secretion of IL-22 by ILC3. Further experiments in vivo can explore the complex mechanisms of antagonist roling in the body. These results are important to provide technique and theory basis for targeted therapy in IBD.

Keywords: Antagonist; RORgt; Th17; ILC3; IBD

1绪论 1

1.1肠道炎症性疾病概述 1

1.2 维甲酸相关孤核受体家族中的RORgt简介 2

1.3 T辅助17细胞及3型天然淋巴细胞简介 3

1.4小分子拮抗剂的简介 5

1.5课题意义 6

2 实验基本原理 8

2.1 T细胞体外诱导 8

2.2肠道固有层淋巴细胞体外刺激 9

2.3 维甲酸相关孤核受体家族中的RORgt拮抗剂的作用 9

2.4流式染色及检验 9

2.5其他重要步骤原理 10

3 实验材料及方法 12

3.1实验原料 12

3.2实验试剂 12

3.3实验主要仪器 13

3.4拮抗剂对T辅助17细胞体外分化影响实验 13

3.5拮抗剂对固有层淋巴细胞体外刺激影响实验 17

4实验结果及讨论小分子化合物作为拮抗剂的筛选及其在肠道炎症性疾病中的应用:http://www.751com.cn/yixue/lunwen_51399.html