核受体RORγt抑制剂的发现及对IL-17A表达影响的研究

Screening of nuclear receptor RORγt inhibitor and its effect on IL-17A expression

目的寻找维甲酸受体相关孤儿受体γt(retinoic receptor-related orphan receptor,RORγt)抑制剂,并对其抗炎活性进行初步研究。方法通过荧光偏振实验(FP assay)对本实验室的化合物库进行高通量筛选,然后再采用荧光素酶报告基因实验(luciferase reporter gene assay)测定目标化合物对RORγt和IL-17在细胞水平的抑制活性,MTT实验检测化合物对细胞毒性的影响。结果通过荧光偏振实验发现了化合物尼麦角林,细胞转染实验结果进一步表明该化合物在细胞水平上对RORγt的转录活性和IL-17A的表达有抑制作用。结论尼麦角林通过抑制RORγt的转录活性,继而影响IL-17A的表达,由此推断该化合物可能具有潜在的抗炎活性。

Retinoic acid-related orphan receptor RORγt is a critical regulator in the differentiation of Th17 cells, which plays a key role in the pathology of several autoimmune diseases. Therefore, RORγt could be apromising target of drug development for IL-17-mediated autoimmune disease. This study designed to seek RORγtinhibitors and evaluate their anti-inflammatory activities preliminarily. A fluorescent polarization assay was used forscreening RORγt ligands which have high affinity to nuclear receptor RORγt. Then cell-based reporter assaysystem was established to screen RORγt inhibitors from high affinity compounds with RORγt. MTT assay wasperformed to determine the cell toxicity of the tested compounds. A compound, Nicergoline, was found as a highbinding RORγt inhibitor which suppressed RORγt activity and IL-17 A expression. In conclusion, Nicergoline,discovered in our study, can inhibit RORγt activity and IL-17 A function, which indicates that Nicergoline could bea potential therapeutic agent against IL-17-derived autoimmune diseases.