生物学杂志

生物学杂志 ›› 2025, Vol. 42 ›› Issue (2): 66-.doi: 10.3969/j.issn.2095-1736.2025.02.066

• 研究报告 • 上一篇    下一篇

葵花盘中黄嘌呤氧化酶抑制活性成分识别与活性分析

闫昊前1,2， 张　扬2， 邢芳毓2， 高建萍2， 张天阳2， 李晓静3, 宋默微4， 孙冰冰3，马　莉1,5， 张贵锋2   

  1. 1. 成都中医药大学 药学院， 成都 610072； 2. 中国科学院过程工程研究所生化工程国家重点实验室，
    北京 100190； 3. 天新福（北京）医疗器材股份有限公司， 北京 102299； 4. 哈尔滨医科大学附属第一医院，
    哈尔滨 150001； 5. 首都医科大学 中医药学院， 北京 100069
  • 出版日期:2025-04-18 发布日期:2025-04-14
  • 通讯作者: 马莉，教授，博士生导师，研究方向为中药质量控制研究，E-mail:marytcm@ccmu.edu.cn；张贵锋，研究员，博士生导师，研究方向为蛋白分析检验研究E-mail:gfzhang@ipe.ac.cn；马莉和张贵锋为共同通信作者
  • 作者简介:闫昊前，硕士研究生，研究方向为中药成分研究，E-mail:yanhaoqian@stu.cdutcm.edu.cn
  • 基金资助:
    北京市自然科学基金项目（2244105）

Identification and characterization of XO-binding natural products from sunflower calathide

YAN Haoqian1,2, ZHANG Yang2, XING Fangyu2, GAO Jianping2, ZHANG Tianyang2,LI Xiaojing3, SONG Mowei4, SUN Bingbing3, MA Li1,5, ZHANG Guifeng2   

  1. 1. School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 610072, China;
    2. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences,
    Beijing 100190, China; 3. Tianxinfu (Beijing) Medical Devices Company Limited, Beijing 102299, China; 4. The First
    Affiliated Hospital of Harbin Medical University, Harbin 150001, China; 5. Schoolc of Traditional Chinese Medicine,
    Capital Medical University, Beijing 100069, China
  • Online:2025-04-18 Published:2025-04-14

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摘要： 从葵花盘中筛选具有黄嘌呤氧化酶（XO）抑制活性的天然产物。以XO为载体，从葵花盘提取物中特异性吸附分离候选化合物，通过高分辨液质谱联用技术对其进行鉴定，研究XO抑制活性成分及其作用机理。从葵花盘醇提物中筛选出咖啡醇等13种化合物，其中，咖啡醇(IC50=47.49 μg/mL，P＜0.0001)具有XO抑制活性，为竞争性可逆抑制剂。采用分子对接技术模拟咖啡醇与XO特异结合性能，通过紫外可见光谱、荧光光谱及圆二色谱技术证明咖啡醇可与XO有效结合，并降低其活性。咖啡醇与酶FAD区域的结合阻碍了底物氧化反应过程中的电子转移，使酶的结构更加致密，底物难以定位到活性腔中，导致其催化活性下降。咖啡醇对酶的猝灭机制属于静态猝灭，主要驱动力为疏水作用，且咖啡醇在XO活性腔上只有单一或一类结合位点。咖啡醇是一种潜在抑制XO活性的成分，为治疗高尿酸血症天然产物的筛选提供依据。

关键词: 葵花盘, 黄嘌呤氧化酶, 咖啡醇, 超滤-质谱联用技术, 尿酸

Abstract: Natural products with inhibition of xanthine oxidase (XO) activity were screened from sunflower calathide. XO was used as a carrier to specifically adsorb and isolate candidate compounds from sunflower calathide extracts, and they were identified by high-resolution liquid mass spectrometry (LC-MS) to study the inhibitory activity and inhibition mechanism of possible compounds on XO. Thirteen compounds were isolated from the ethanol extract of sunflower calathide, including cafestol (IC50=47.49 μg/mL,P<0.0001), which exhibited competitive and reversible inhibition against XO. Circular dichroism with molecular docking verified that cafestol could bind to XO and interact with it and reduce its activity. The binding of cafestol to the FAD region of the enzyme hindered the electron transfer during the substrate oxidation reaction, making the structure of the enzyme more dense, and the substrate was difficult to locate into the active cavity, resulting in a decrease in its catalytic activity. The quenching mechanism of cafestol against xanthine oxidase belonged to static quenching, and the main driving force was hydrophobic, and cafestol had only a single or one binding site on the XO active cavity. The results showed that cafestol was a potential ingredient that inhibited XO activity, which provided a scientific basis for the screening of therapeutic drugs such as hyperuricemia.

Key words: sunflower calathide, XO, cafestol, UF-LC-MS, uric acid

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