基于LC/Q-TOF-MS整合网络药理学探讨潜阳育阴颗粒防治高血压肾损害的药效物质基础与作用机制  

Investigation of Pharmacodynamic Material Basis and Mechanism of Qianyang Yuyin Granules Against Hypertensive Renal Damage Based on LC/Q-TOF-MS and Network Pharmacology

作　　者：吴信华 李长印[1] 曹园[1] 瞿慧[1] 韩洋 方祝元[1] WU Xin-hua;LI Chang-yin;CAO Yuan;QU Hui;HAN Yang;FANG Zhu-yuan(Affiliated Hospital of Nanjing University of Chinese Medicine,Nanjing 210029,China)

机构地区：[1]南京中医药大学附属医院,南京210029

出　　处：《中国实验方剂学杂志》2021年第2期116-129,共14页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：江苏省中医药局科技项目(JD201813);南京中医药大学优势学科项目(ZYX03KF074)。

摘　　要：目的:系统研究潜阳育阴颗粒的化学成分,探讨其发挥药效的主要活性成分和作用机制。方法:采用液相色谱/四级杆-飞行时间质谱法(LC/Q-TOF-MS)对潜阳育阴颗粒中的化学成分进行全面分析,检测条件为Agilent Poroshell 120 SBC18色谱柱(3.0 mm×100 mm,2.7μm),流速0.4 mL·min^(-1),电喷雾离子源(ESI),正、负离子模式扫描,扫描范围m/z 25~1 000;正离子模式的流动相A为水+10 mmol·L^(-1)甲酸铵+0.125%甲酸+0.1%甲醇,B为乙腈-水(9∶1)+10 mmol·L^(-1)甲酸铵+0.125%甲酸;负离子模式的流动相A为水+10 mmol·L^(-1)甲酸铵+0.1%甲醇,B为乙腈-水(9∶1)+10 mmol·L^(-1)甲酸铵;梯度洗脱程序均设定为0~3.5 min,5%B;3.5~4 min,5%~10%B;4~9 min,10%~25%B;9~18 min,25%~30%B;18~25 min,30%~50%B;25~27 min,50%~90%B;27~32 min,90%B;32~33 min,90%~5%B;33~39 min,5%B。根据化合物的一级、二级质谱信息,与对照品或文献数据比对,快速识别化学成分。结合网络药理学方法对潜阳育阴颗粒的化学成分进行靶标分析、功能富集,初步筛选出主要药效物质,并探讨其作用机制。结果:从潜阳育阴颗粒中共识别出99个化学成分,从中筛选出43个与高血压肾损害相关的活性成分及48个核心靶点。核心成分包括山柰酚,槲皮素,阿魏酸,木樨草素,咖啡酸甲酯,肉桂酸,芦荟大黄素,大黄素,N-反式-阿魏酰酪胺,没食子酸,异红草素,8-O-阿魏酰基哈巴苷,咖啡酸乙酯,异奥卡宁,杯苋甾酮,二苯乙烯苷,马钱苷,23-乙酰泽泻醇B和哈巴苷;核心靶点包括血管内皮生长因子A(VEGFA),丝氨酸/苏氨酸蛋白激酶1(AKT1),Jun原癌基因(JUN),G蛋白β1(GNB1),肿瘤蛋白p53(TP53),β-淀粉样前体蛋白(APP),丝裂原活化蛋白激酶1(MAPK1)等。结论:潜阳育阴颗粒主要从调控血管内皮、血管生成、炎症反应、氧化应激及免疫应答等方面发挥清肝补肾、化瘀通络的功效。Objective: To systematically study the chemical components of Qianyang Yuyin granules and explore its main pharmacodynamic substances and mechanism in the prevention and treatment of hypertensive renal damage. Method: Liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(LC/Q-TOF-MS) was employed to comprehensively analyze the chemical components of Qianyang Yuyin granules. Agilent Poroshell 120 SB-C18 column(3.0 mm×100 mm,2.7 μm)was used,flow rate was 0.4 mL·min^(-1),electrospray ionization(ESI)was applied and operated in positive and negative ion modes,the acquisition range was m/z 25-1 000. Mobile phase in positive ion mode consisted of water+10 mmol·L^(-1) ammonium formate+0.125% formic acid+0.1% methanol(A)-[acetonitrile-water(9∶ 1)+10 mmol·L^(-1) ammonium formate+0.125% formic acid](B),and in negative ion mode consisted of water+10 mmol·L^(-1) ammonium formate+0.1% methanol(A)-[acetonitrile-water(9∶1)+10 mmol·L^(-1) ammonium formate](B)with the gradient elution(0-3.5 min,5%B;3.5-4 min,5%-10%B;4-9 min,10%-25%B;9-18 min,25%-30%B;18-25 min,30%-50%B;25-27 min,50%-90%B;27-32 min,90%B;32-33 min,90%-5%B;33-39 min,5%B). According to the information of the accurate mass,the multistage fragment ions,the mass spectrometric data of the standard substances and the relative reference literature,the structures of the chemical components in Qianyang Yuyin granules were identified. Based on the identified components,network pharmacology study,including target prediction and functional enrichment was applied to screen out the main active substances against hypertensive renal damage,and explore the potential mechanism. Result: A total of 99 chemical components were identified,from which 43 active substances and 48 key targets were screened out. The key components contained kaempferol,quercetin,ferulic acid,luteolin,caffeic acid methyl ester,cinnamic acid,aloe-emodin,emodin,gallic acid,N-trans-feruloyltyramine,isoorientin,8-O-feruloylharpagide,ethyl caffeate,isookanin,cyasterone, 2,3,5,4’-

关 键 词：潜阳育阴颗粒 高血压肾损害 网络药理学 液相色谱/四级杆-飞行时间质谱法(LC/Q-TOF-MS) 药效物质 作用机制 数据库 

分 类 号：R22[医药卫生—中医基础理论] R96[医药卫生—中医学]