基于UPLC-Q-TOF-MS联合网络药理学探讨驱虫斑鸠菊注射液诱导黑色素积累的作用机制  

Melanogenesis of quality markers in Vernonia anthelmintica Injection based on UPLC-Q-TOF-MS combined network pharmacology

作　　者：罗林 张晏源 王程 黄思露 王晓琴 张波[1,2] LUO Lin;ZHANG Yan-yuan;WANG Cheng;HUANG Si-lu;WANG Xiao-qin;ZHANG Bo(Key Laboratory of Xinjiang Phytomedicine Resource and Utilization,Ministry of Education,School of Pharmacy,Shihezi University,Shihezi 832002,China;Sichuan Industrial Institute of Antibiotics,School of Pharmacy,Chengdu University,Chengdu 610106,China)

机构地区：[1]新疆植物药资源利用教育部重点实验室,石河子大学药学院,新疆石河子832002 [2]四川抗菌素工业研究所,成都大学药学院,四川成都610106

出　　处：《中国中药杂志》2023年第6期1606-1619,共14页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(U1603122);新疆生产建设兵团创新领域中青年领军人才项目(2018CB019)。

摘　　要：评价驱虫斑鸠菊注射液(VAI)促黑色素积累的效应机制。建立丙基硫氧嘧啶(PTU)体内脱色素斑马鱼模型,结合B16F10细胞体外模型评价VAI诱导黑色素积累效应。通过UPLC-Q-TOF-MS技术鉴定VAI化学成分,采取网络药理学方法预测VAI的潜在靶点与通路,建立“VAI成分-靶点-通路”网络图,基于拓扑学特征筛选药效分子。分子对接验证药效分子与关键靶点的结合。结果显示VAI呈剂量和时间依赖性促进B16F10细胞酪氨酸酶活性和黑色素生成,且能恢复上述斑马鱼模型体内黑色素。从VAI中共鉴定出56种化合物,其中有黄酮类(15/56)、萜类(10/56)、酚酸类(9/56)、脂肪酸类(9/56)、甾体类(6/56)和其他(7/56)。网络药理学手段分析筛选出芹菜素、金圣草黄素、丁香脂素、紫铆花素4个潜在质量标志物,涉及61个靶点和65条通路,分子对接验证了其与TYR、NFE2L2、CASP3、MAPK1、MAPK8、MAPK14结合并发现VAI能促进B16F10细胞内MITF、TYR、TYRP1、DCT等基因mRNA的表达。该文通过UPLC-Q-TOF-MS和网络药理学技术确定了VAI抗白癜风的物质基础,优选出芹菜素、金圣草黄素、丁香脂素、紫铆花素为其质量标志物,验证了其促黑色素生成的药效和内在机制,为质量控制和进一步临床研究提供依据。This study aimed to evaluate the biological effect and mechanism of Vernonia anthelmintica Injection(VAI)on melanin accumulation.The in vivo depigmentation model was induced by propylthiouracil(PTU)in zebrafish,and the effect of VAI on melanin accumulation was evaluated based on the in vitro B16F10 cell model.The chemical composition of VAI was identified according to the high-performance liquid chromatography quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS).Network pharmaco-logy was applied to predict potential targets and pathways of VAI.A"VAI component-target-pathway"network was established,and the pharmacodynamic molecules were screened out based on the topological characteristics of the network.The binding of active molecules to key targets was verified by molecular docking.The results showed that VAI promoted tyrosinase activity and melanin production in B16F10 cells in a dose-and time-dependent manner and could restore the melanin in the body of the zebrafish model.Fifty-six compounds were identified from VAI,including flavonoids(15/56),terpenoids(10/56),phenolic acids(9/56),fatty acids(9/56),steroids(6/56),and others(7/56).Network pharmacological analysis screened four potential quality markers,including apigenin,chrysoeriol,syringaresinol,and butein,involving 61 targets and 65 pathways,and molecular docking verified their binding to TYR,NFE2L2,CASP3,MAPK1,MAPK8,and MAPK14.It was found that the mRNA expression of MITF,TYR,TYRP1,and DCT in B16F10 cells was promoted.By UPLC-Q-TOF-MS and network pharmacology,this study determined the material basis of VAI against vitiligo,screened apigenin,chrysoeriol,syringaresinol,and butein as the quality markers of VAI,and verified the efficacy and internal mechanism of melanogenesis,providing a basis for quality control and further clinical research.

关 键 词：驱虫斑鸠菊注射液 质量标志物 网络药理学 UPLC-Q-TOF-MS 黑色素生成 

分 类 号：R285[医药卫生—中药学]