基于超高效液相色谱/四极杆-静电场轨道阱质谱联合网络药理学筛选“芩百清肺浓缩丸”治疗肺炎的活性成分  

Screening of the Active Components of Qinbai Qingfei Concentrated Pill on Treating Pneumonia Based on UHPLC/Q-Exactive Orbitrap MS Uniting Network Pharmacology

作　　者：于洋 辛杨 王海军[3] 杨清竹[4] 王伟明 YU Yang;XIN Yang;WANG Hai-jun;YANG Qing-zhu;WANG Wei-ming(Heilongjiang Academy of Chinese Medical Sciences,Harbin 150036,China;College of Chemistry and Chemical Engineering,Qiqihar University,Qiqihar 161006,China;School of General Medicine and Continuing Education,Qiqihar Medical University,Qiqihar 161006,China;College of Life Science and Agroforestry,Qiqihar University,Qiqihar 161006,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036 [2]齐齐哈尔大学化学与化学工程学院,黑龙江齐齐哈尔161006 [3]齐齐哈尔医学院全科医学与继续教育学院,黑龙江齐齐哈尔161006 [4]齐齐哈尔大学生命科学与农林学院,黑龙江齐齐哈尔161006

出　　处：《质谱学报》2024年第6期884-896,I0006,共14页Journal of Chinese Mass Spectrometry Society

基　　金：国家自然科学基金(82174060);黑龙江省省属高等学校基本科研业务费科研项目(145209505);黑龙江省博士后资助项目(LBH-Z19099)。

摘　　要：为筛选中药地龙在芩百清肺浓缩丸治疗肺炎时贡献的活性成分,本研究在开展细胞实验证实芩百清肺浓缩丸抗炎作用的基础上,采用超高效液相色谱/四极杆-静电场轨道阱质谱(UHPLC/Q-Exactive Orbitrap MS)法检测有、无地龙的芩百清肺浓缩丸提取液,借助Compound Discoverer软件筛选差异离子,根据二级质谱数据及标准品鉴定差异成分,进一步开展差异成分治疗肺炎的网络药理学研究,预测活性成分与靶点,并通过分子对接与实时荧光定量逆转录聚合酶链反应(RT-qPCR)技术验证活性成分与靶点。结果表明,芩百清肺浓缩丸可通过降低细胞炎症因子IL6、IL-1b,趋化因子CXCL2和CXCR2的表达发挥抗炎活性,通过液相色谱-质谱筛选得到了15个差异成分,网络药理预测得到7个潜在活性成分及22个核心靶点,其中α-亚麻酸和腺苷经分子对接及RTqPCR实验验证为地龙在芩百清肺浓缩丸治疗肺炎时贡献的活性成分。该方法为筛选中药复方中组方药材贡献活性成分提供了参考,为中药复方活性成分研究提供了思路。In order to screen the active ingredients of Qinbai Qingfei Concentrated Pill(QQCP)on treating pneumonia contributed by Pheretima,the extracts of QQCP with and without Pheretima were detected by ultra-high performance liquid chromatography/quadrupole-exactive Orbitrap mass spectrometry(UHPLC/Q-Exactive Orbitrap MS)after conducting the cell anti-inflammatory experiment.Compound Discoverer(CD)software was used to screen the differential ions,and the differential components were identified according to MS2 spectra and standards.Further,the active ingredients and targets were predicted by carrying out network pharmacology of differential components treating pneumonia.At last,active ingredients which were identified by standards and their corresponding targets were verified by molecular docking and real-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-qPCR).The results of cell anti-inflammatory experiments showed that QQCP can exert anti-inflammatory activity by decreasing the expression of IL6,IL-1b,CXCL2 and CXCR2.After detecting the extracts of QQCP by applying UHPLC/Q-Exactive Orbitrap MS,a total of 15 components are identified on the basis of their retention time,MS/MS spectra as well as standards.After that,there are 7 components connecting and 22 core targets obtained by applying networkpharmacology.After verified by molecular docking and RT-qPCR,α-linolenic acid and adenosine are considered as the active components of QQCP treating pneumonia.This study develops a method of UHPLC/Q-Exactive Orbitrap MS uniting network pharmacology to screen the contributing active components from one medicine in Chinese medicine compounds,which provides a new idea for the study on active components of Chinese medicine compounds.

关 键 词：超高效液相色谱/四极杆-静电场轨道阱质谱(UHPLC/Q-Exactive Orbitrap MS) 网络药理 芩百清肺浓缩丸 α-亚麻酸 腺苷 肺炎 

分 类 号：O657.63[理学—分析化学]