基于斑马鱼模型及分子对接技术探究艾叶黄酮的降尿酸作用及药效活性成分  

Investigation of uric acid-lowering effect and bioactive components of flavonoids from Artemisiae Argyi Folium based on zebrafish model and molecular docking techniques

作　　者：刘勇杰 王平 夏婧 朱澳华 昝俊峰 陈林霖 刘军锋 LIU Yongjie;WANG Ping;XIA Jing;ZHU Aohua;ZAN Junfeng;CHEN Linlin;LIU Junfeng(College of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Institute of Gerontology,Hubei University of Chinese Medicine,Wuhan 430065,China;Key Laboratory of Traditional Chinese Medicine Resources and Compounds,Ministry of Education,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]湖北中医药大学老年医学研究所,湖北武汉430065 [3]湖北中医药大学中药资源与中药复方教育部重点实验室,湖北武汉430065 [4]湖北时珍实验室,湖北武汉430065

出　　处：《中草药》2024年第24期8470-8478,共9页Chinese Traditional and Herbal Drugs

基　　金：湖北省自然科学基金-中医药创新发展联合基金资助项目(2024AFD285)。

摘　　要：目的探索艾叶Artemisiae Argyi Folium黄酮的降尿酸(uric acid,UA)作用及发挥药效的活性成分。方法采用氧嗪酸钾(250μmol/L)联合黄嘌呤(10μmol/L)建立急性高尿酸血症斑马鱼模型,艾叶醇提物(250、500、1000μg/mL)处理后检测斑马鱼体内UA、丙二醛(malondialdehyde,MDA)、活性氧(reactive oxygen species,ROS)水平及超氧化物歧化酶(superoxide dismutase,SOD)和黄嘌呤氧化酶(xanthine oxidase,XO)活性。通过艾叶黄酮与XO的分子对接,分析各黄酮类化合物与XO的结合模式并计算比较结合能。通过XO抑制率实验研究艾叶黄酮对XO的抑制率。结果艾叶醇提物能显著降低高尿酸血症模型斑马鱼UA、MDA、ROS水平(P<0.05、0.01、0.001),显著升高SOD活性(P<0.05、0.01、0.001),显著降低XO活性(P<0.01)。分子对接结果显示,多数艾叶黄酮化合物均能与XO关键氨基酸结合,且与XO的结合能较低。XO抑制率实验结果表明木犀草素、槲皮素、山柰酚、泽兰黄酮和芹菜素能明显抑制XO活性。结论艾叶醇提物具有明显的降UA作用,其机制可能是黄酮类化合物抑制XO活性,其主要发挥降UA作用的化合物可能是木犀草素、槲皮素、山柰酚、泽兰黄酮和芹菜素。Objective To explores the uric acid(UA)-lowering effects of flavonoids from Aiye(Artemisiae Argyi Folium)and their bioactive constituents responsible for pharmacological activity.Methods An acute hyperuricemia zebrafish model was established using oxonic acid potassium(250μmol/L)combined with xanthine(10μmol/L).The zebrafish were treated with ethanol extracts of Artemisiae Argyi Folium(250,500,1000μg/mL),and the levels of UA,malondialdehyde(MDA),reactive oxygen species(ROS),superoxide dismutase(SOD)activity,and xanthine oxidase(XO)activity were measured.Molecular docking was used to analyze the binding modes between Artemisiae Argyi Folium flavonoids and XO and calculate binding energies.The inhibition rates of flavonoids on XO were studied through XO inhibition assays.Results The ethanol extract of Artemisiae Argyi Folium significantly reduced UA,MDA and ROS levels in the hyperuricemia zebrafish model(P<0.05,0.01,0.001),while significantly increasing SOD activity(P<0.05,0.01,0.001)and significantly reducing XO activity(P<0.01).Molecular docking results showed that most flavonoids from Artemisiae Argyi Folium could bind to key amino acids of XO with relatively low binding energies.XO inhibition assays indicated that luteolin,quercetin,kaempferol,eupatilin,and apigenin could significantly inhibit XO activity.Conclusion The ethanol extract of Artemisiae Argyi Folium has a significant UA-lowering effect,likely due to the inhibition of XO activity by its flavonoid compounds.The main compounds contributing to the UA-lowering effect may be luteolin,quercetin,kaempferol,nepetin,and apigenin.

关 键 词：艾叶黄酮 黄嘌呤氧化酶 降尿酸 分子对接 斑马鱼 木犀草素 泽兰黄酮 芹菜素 

分 类 号：R285.5[医药卫生—中药学]