基于UPLC-Q-TOF/MS整合网络药理学探究红景天口服液抗急性高原反应的活性成分及作用机制  

Exploring the active components and mechanism of anti-acute mountain sickness of Hongjingtian oral liquid based on UHPLC-QTOF/MS and network pharmacology

作　　者：陈奎奎 闫晓宁 郭忠会 沈琳 梁洁 冯玉燕[1] 肖红斌 CHEN Kuikui;YAN Xiaoning;GUO Zhonghui;SHEN Lin;LIANG Jie;FENG Yuyan;XIAO Hongbin(School of Pharmacy,Guangxi University of Traditional Chinese Medicine,Nanning 530200,China;Research Center for Chinese Medicine Analysis and Transformation,Beijing University of Chinese Medicine,Beijing 100029,China;Guangxi Zhuang Yao Key Laboratory of Medicine,Guangxi University of Chinese Medicine,Nanning 530200,China;Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]广西中医药大学药学院,广西南宁530200 [2]北京中医药大学中药分析与转化研究中心,北京100029 [3]广西中医药大学广西壮瑶药重点实验室,广西南宁530200 [4]北京中医药大学北京中医药研究院,北京100029

出　　处：《沈阳药科大学学报》2024年第12期1531-1545,共15页Journal of Shenyang Pharmaceutical University

基　　金：广西中医药大学青年基金项目(2021QN006);广西青年科学基金(2023GXNSFBA026276);广西科学技术厅广西科技基地与人才专项(桂科AD21238032);科学技术部重大新药创制国家科技重大专项课题(2019ZX09201004);广西中医药大学第三批“岐黄工程”高层次人才团队培育项目(202406)。

摘　　要：目的探究红景天口服液的活性成分及其抗急性高原反应的机制。方法采用超高效液相色谱-四级杆飞行时间串联质谱(ultra-performance liquid chromatography-quadrupole time of flight mass spectrometry,UPLC-Q-TOF/MS)法表征红景天口服液的主要化学成分。运用网络药理学方法检索红景天口服液和急性高原反应(acute mountain sickness,AMS)的潜在靶点,将药物靶点与AMS相关靶点取交集,基于STRING数据库进行靶点蛋白相互作用网络(protein-protein interac-tion,PPI)分析,借助DAVID数据库平台进行基因本体(gene ontology,GO)功能分析及京都基因与基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)通路富集分析。考察红景天苷、红景天素、对羟基苯甲醇、酪醇4个活性成分对糖氧剥夺诱导嗜铬细胞瘤12(pheochromocytoma 12,PC12)细胞损伤模型的存活率及乳酸脱氢酶(lactate dehydrogenase,LDH)露出率的影响。结果从红景天口服液中鉴定了40个主要的化学成分,进一步筛选出抗AMS活性成分14个,所对应相关靶点34个,其中肿瘤坏死因子(tumor necrosis factor,TNF)、白介素6(interleukin 6,IL-6)、血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)、环加氧酶2(prostaglandin-endoper-oxide synthase 2,PTGS2)等靶点的关联性较强;红景天口服液治疗AMS主要涉及磷脂酰肌醇3-激酶-蛋白激酶B(phosphatidylinositol 3-kinase-protein kinase B,PI3K-Akt)、缺氧诱导因子1(hypoxi-a-inducible factor 1,HIF-1)和丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)等信号通路。红景天苷、红景天素、对羟基苯甲醇、酪醇可显著提高糖氧剥夺PC12细胞损伤模型的存活率及减少其LDH露出率。结论红景天口服液中的红景天苷、酪醇、红景天素、对羟基苯甲醇等可能是抗急性高原反应的潜在药效物质基础,作用机制可能与HIF-1、MAPK、PI3K-Akt等信号通路有关。Objective To explore the active ingredients of Hongjingtian Oral Liquid and the mechanism against acute mountain sickness.Methods Ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF/MS)was applied to profile the main chemical constituents of Hongjingtian Oral Liquid.Potential targets of Hongjingtian Oral Liquid and Acute mountain sickness(AMS)were retrieved by network pharmacological methods,drug targets were intersected with AMS-related targets.The target protein-protein interaction network(PPI)was analyzed based on the STRING database.With the help of DAVID database,the gene ontology(GO)function analysis and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were performed.The effects of salidroside,rhodiosin,p-hydroxybenzyl alcohol and tyrosol on the survival rate and lactate dehydrogenase(LDH)exposure rate of Pheochromocytoma 12(PC12)cell injury model induced by glucose and oxygen deprivation were investigated.Results Forty main chemical components were identified from Hongjingtian Oral Liquid,and 14 anti-AMS active components were screened,and 34 related targets were identified.Among them,tumor necrosis factor(TNF),interleukin-6(IL-6),vascular endothelial growth factor A(VEGFA)and prostaglandin-endoperoxide synthase 2(PTGS2)were highly correlated.The treatment for AMS of Hongjingtian Oral Liquid mainly involves signaling pathways such as phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt),hypoxia-inducible factor 1(HIF-1)and mitogen-activated protein kinase(MAPK),etc.Salidroside,rhodiosin,p-hydroxybenzyl alcohol and tyrosol can improve the survival rate of PC12 cell injury model and reduce the exposure rate of LDH.Conclusion The salidroside,tyrosol,rhodiosin and p-hydroxybenzyl alcohol in Hongjingtian Oral Liquid may be the potential pharmacodynamic material basis,and may play an anti-AMS role by regulating HIF-1,MAPK,PI3K-Akt and other signaling pathways.

关 键 词：红景天口服液 急性高原反应 UPLC-Q-TOF/MS 网络药理学 活性成分 作用机制 

分 类 号：R285[医药卫生—中药学]