复方芩兰口服液的体内外成分分析和网络药理学研究

In vivo and in vitro component analysis and network pharmacology study of Fufang Qinlan Oral Liquid

目的采用液质联用技术对复方芩兰口服液的主要化学成分及其进入血液和组织中的成分进行鉴定,并选择主要的体内成分开展网络药理学研究,初步揭示其作用机制。方法采用UPLC-Q-TOF-MS结合MSE技术进行复方芩兰口服液、血液和组织样本分析,结合精确相对分子质量、保留时间、碎片离子、中性丢失等信息进行复方芩兰口服液的化学成分和体内成分鉴定。利用Cytoscape软件分别构建复方芩兰口服液主要体内成分与其功能主治(感冒、发热、咳嗽、咽痛)之间的“化合物-靶点”作用网络,开展核心靶点和作用通路分析,并采用实时荧光定量PCR对核心靶点中的3个热休克蛋白(HSP90AA1、HSP90AB1、HSPA8)进行生物学验证。结果大鼠ig给药后在血液样本中检测到3个原型成分和9个代谢产物,组织样本中检测到3个原型成分和5个代谢产物,代谢产物主要为葡萄糖醛酸结合物。网络药理学研究结果提示复方芩兰口服液的作用通路主要与炎症和免疫相关,且生物学验证结果表明上呼吸道感染大鼠肺脏组织中HSP90AA1、HSP90AB1、HSPA8的m RNA表达在给予复方芩兰口服液后显著上调(P<0.01)。结论利用UPLC-Q-TOF-MS分析复方芩兰口服液的体内外成分并进行网络药理学研究,初步明确了复方芩兰口服液的作用靶点和通路,并选择3个关键靶点进行生物学验证。研究结果为进一步揭示其药效物质基础和作用机制提供科学依据。

Objective Liquid mass spectrometry was used to identify the chemical constituents of Fufang Qinlan Oral Liquid(复方芩兰口服液) and its metabolites in rat.The main in vivo components were selected for network pharmacological and the mechanism was initially revealed.Methods Fufang Qinlan Oral Liquid,serum and tissue samples were analyzed by UPLC-Q-TOF-MS combined with MSE technique,and the identification of in vivo and in vitro components were carried out with the information of precise molecular weight,retention time,fragment ions and neutral loss.Cytoscape software was used to construct the “compound-target” network between the main in vivo components and the main clinical indications(cold,fever,cough and sore throat).Core targets and pathway analysis were also conducted,in which HSP90AA1,HSP90AB1 and HSPA8 were measured by realtime PCR.Results Three prototype components and nine metabolites were characterized in rat serum after ig administration,so as three prototype components and five metabolites in rat tissues.The metabolites were mainly glucuronide conjugates.The results of network pharmacology indicated that the pathways of Fufang Qinlan Oral Liquid were mainly related to inflammation and immunity.Compared with model rats,the expression of HSP90AA1,HSP90AB1,and HSPA8 was significantly increased after intragastric administration of Fufang Qinlan Oral Liquid(P<0.01).Conclusion The in vivo components were preliminarily analyzed after oral administration of Fufang Qinlan Oral Liquid by UPLC-Q-TOF-MS and the network pharmacology study was then carried out,which initially clarified the targets and pathways.The studies provided scientific basis for pharmacodynamic material basis and mechanism of Fufang Qinlan Oral Liquid.