UPLC-Q-Orbitrap HRMS结合分子对接探究肉苁蓉苦味的物质基础及味-效关系  

Material basis of bitter taste and taste-effect relationship in Cistanche deserticola based on UPLC-Q-Orbitrap HRMS combined with molecular docking

作　　者：田利颖 李明杰 侯强 王郑园 古力孜也·艾赛提 胡君萍[1] TIAN Li-ying;LI Ming-jie;HOU Qiang;WANG Zheng-yuan;CULIZIYE Ai-sai-ti;HU Jun-ping(College of Pharmacy,Xinjiang Medical University,Urumqi 830017,China;Department of Pharmacy,Shaya People's Hospital,Shaya 842200,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830017 [2]沙雅县人民医院药剂科,新疆沙雅842200

出　　处：《中国中药杂志》2025年第6期1569-1580,共12页China Journal of Chinese Materia Medica

基　　金：新疆维吾尔自治区自然科学基金重点项目(2021D01D11,2022D01D14);新疆维吾尔自治区自然科学基金地州科学基金项目(2022D01F92);新疆天然药物有效成分与药物释放技术重点实验室项目(XJDX1713)。

摘　　要：基于超高效液相色谱-四极杆-静电场轨道阱高分辨质谱(UPLC-Q-Orbitrap HRMS)技术和分子对接技术探究肉苁蓉提取物的苦味呈味物质(以下简称呈苦物质),并探究其与苦味功效的关系,为脱苦及矫味奠定研究基础。首先采用UPLC-Q-Orbitrap HRMS对肉苁蓉成分进行定性分析,鉴定出69个化学成分,将这些化学成分与苦味受体进行分子对接,筛选出20个呈苦物质,包括6个苯乙醇苷类、5个黄酮类、3个酚酸类、2个环烯醚萜类、2个生物碱类、2个其他类成分。采集相同产地不同月份的9批次新鲜肉苁蓉样品,将其分为不同月份及不同部位,采用电子舌法测定样品的苦味量化值,采用高效液相色谱法(HPLC)测定样品中6个潜在苦味药效成分(松果菊苷、毛蕊花糖苷、管花苷A、异毛蕊花糖苷、金石蚕苷、京尼平苷)的含量,采用紫外-可见分光光度法测定样品中总苯乙醇苷、总多糖、总生物碱、总黄酮、总酚酸含量,并将在此基础上结合化学计量学分析,通过皮尔逊(Pearson)相关性分析、灰色关联分析、正交偏最小二乘法判别分析寻找肉苁蓉中的苦味成分,均与分子对接结果一致,2种方法互为佐证。最后以呈苦物质为研究对象,进行网络药理学预测与分析,并探究了呈苦物质作用靶点与苦味功效的关系,结果表明呈苦物质作用的关键靶点为EGFR、PIK3CB、PTK2等,并且呈苦物质可能通过作用于相关疾病靶点而发挥苦味功效,即肉苁蓉呈苦物质是苦味功效的物质基础。综上,该研究表明以松果菊苷、毛蕊花糖苷、金石蚕苷为主的苯乙醇苷类成分为肉苁蓉“苦味”的物质基础,分子对接技术对中药呈苦物质的筛选具有指导作用,肉苁蓉的呈苦物质具有苦味功效,为今后中药的“味-效”关系研究提供思路与参考。Based on ultra-performance liquid chromatography-quadrupole-electrostatic field Orbitrap high-resolution mass spectrometry(UPLC-Q-Orbitrap HRMS)technology and molecular docking,the bitter-tasting substances(hereafter referred to as"bitter substances")in Cistanche deserticola extract were investigated,and the bitter taste and efficacy relationship was explored to lay the foundation for future research on de-bittering and taste correction.Firstly,UPLC-Q-Orbitrap HRMS was used for the qualitative analysis of the constituents of C.deserticola,and 69 chemical components were identified.These chemical components were then subjected to molecular docking with the bitter taste receptor,leading to the screening of 20 bitter substances,including 6 phenylethanol glycosides,5 flavonoids,3 phenolic acids,2 cycloalkenyl ether terpenes,2 alkaloids,and 2 other components.Nine batches of fresh C.deserticola samples were collected from the same origin but harvested at different months.These samples were divided into groups based on harvest month and plant part.The bitterness was quantified using an electronic tongue,and the content of six potential bitter-active compounds(pineconotyloside,trichothecene glycoside,tubulin A,iso-trichothecene glycoside,jinshihuaoside,and jingnipinoside)was determined by high-performance liquid chromatography(HPLC).The total content of phenylethanol glycosides,polysaccharides,alkaloids,flavonoids,and phenolic acids was determined using UV-visible spectrophotometry.Chemometric analyses were then conducted,including Pearson′s correlation analysis,gray correlation analysis,and orthogonal partial least squares discriminant analysis(OPLS-DA),to identify the bitter components in C.deserticola.The results were consistent with the molecular docking findings,and the two methods mutually supported each other.Finally,network pharmacological predictions and analyses were performed to explore the relationship between the targets of bitter substances and their efficacy.The results indicated that key targets of the

关 键 词：肉苁蓉提取物 UPLC-Q-Orbitrap HRMS 网络药理学 分子对接 电子舌 呈苦物质 “味-效”关系 

分 类 号：R284.1[医药卫生—中药学]