【作者】 刘涛；

【导师】 王一飞；

【作者基本信息】 暨南大学 ， 工科 生物工程（专业学位）， 2022， 硕士

【摘要】 目的:为了进一步从北艾中发现新的抗炎活性分子,丰富北艾抗炎活性物质的发现,本研究有目的性的从北艾中富集分离倍半萜类化合物,并进行抗炎活性研究,为北艾抗炎药物研发提供依据。方法:以TLC为指导从北艾95%乙醇提取物中富集倍半萜类化合物,利用多种色谱层析技术分离单体化合物,利用波谱和光谱学技术对单体化合物进行结构与构型的测定,通过MTT实验测试了化合物的细胞毒性,利用Griese法测式化合物的抗炎活性,最后利用生物信息学技术对新化合物进行抗炎活性靶点预测。结果:对42 kg北艾叶进行分离提取,从中分离得到倍半萜类化合物26个,脂肪族类化合物2个,苯系衍生物2个,其中7个为倍半萜类新化合物。抗炎研究表明,愈创木内酯具有较好的抗炎活性,IC₅₀为0.9-4.0 μM,化合物artemvulactone E具有良好抗炎活性,其对LPS诱导的NO半数抑制浓度IC₅₀为0.9 μM,artemvulactone E呈剂量依赖性降低LPS诱导的COX-2蛋白的表达,分子对接预测其抗炎靶点为iNOS和COX-2。结论:本研究从北艾中分离鉴定26个倍半萜类化合物,其中7个为新化合物,分别为artemvulactone A-F和artemvulemdiol A,其余倍半萜类化合物是首次从北艾中分离鉴定。Artemvulactone E具有更良好的抗炎活性,其抑制NO的IC₅₀为0.9 μM,且呈剂量依赖性降低LPS诱导的COX-2蛋白的表达,其可作为抗炎候选物进行进一步药理机制研究。更多还原

【Abstract】 Objective:In order to further discover new anti-inflammatory active molecules and enrich the discovery of anti-inflammatory active substances from Artemisia vulgaris L.This study purposefully isolated sesquiterpenoids and conducted anti-inflammatory activity studies from Artemisia vulgaris L.Methods:Using TLC as a guide to enrich sesquiterpenoids from Artemisia vulgaris,use various chromatographic techniques to separate monomer compounds,use spectral and spectroscopic techniques to determine the structure and configuration of compounds.The cytotoxicity of compounds was examined by MTT assay,the anti-inflammatory activity of compounds was evaluated by Griese assay in a model of LPS-induced inflammation in RAW 264.7 cells,and the anti-inflammatory activity targets of the new compounds were predicted by bioinformatics technology.Results:Twenty-six sesquiterpenoids,two aliphatic compounds and two benzene derivatives were isolated from the leaves of 42 kg Artemisia vulgaris L.Guaiacene sesquiterpenes lactones has obvious anti-inflammatory activity with IC₅₀ of 0.9±4.0μM.Artemvulactone E has significant anti-inflammatory activity with IC₅₀ of 0.9 μM,which decreased LPS-induced COX-2 protein expression in a dose-dependent manner and its anti-inflammatory target is predicted to be iNOS and COX-2 by molecular docking.Conclusion:In this study,Twenty-six sesquiterpenoids including seven undescribed sesquiterpenoids named artemvulactone A-F and artemvulemdiol A were isolated and identified from Artemisia vulgaris L.The remaining sesquiterpenoids were isolated and identified from Artemisia vulgaris L.for the first time.Anti-inflammatory activity experiments show that artemvulactone E has better anti-inflammatory activity,the NO inhibitory activity IC₅₀ value of 0.9 μM,and it can reduce LPS-induced COX-2 protein expression in a dose-dependent manner.Artemvulactone E can be used as an anti-inflammatory candidate for further pharmacological mechanism studies.更多还原