Analysis of Q-markers of Arisaema Cum Bile Acting on Stroke Based on UPLC-QTOF-MS/MS and TCMIP v2.0 / 中国实验方剂学杂志

ABSTRACT

ObjectiveTo predict the possible quality markers （Q-markers） of Arisaema Cum Bile in the prevention and treatment of stroke based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry （UPLC-QTOF-MS/MS） and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine （TCMIP） v2.0. MethodUPLC-QTOF-MS/MS was employed with the mobile phase of 0.1% formic acid aqueous solution （A）-0.1% formic acid acetonitrile solution （B） for gradient elution （0-3 min， 0.2%-5%B； 3-5 min， 5%-8%B； 5-8 min， 8%-10%B； 8-14 min， 10%-25%B； 14-18 min， 25%-50%B； 18-20 min， 50%-70%B； 20-21 min， 70%-98%B； 21-23 min， 98%B； 23-24 min， 98%-0.2%B； 24-26 min， 0.2%B）， the flow rate of 0.5 mL·min-1 and electrospray ionization （ESI）. High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. A local database of the chemical constituents in Arisaema Cum Bile was established by UNIFI 1.8. Then the chemical constituents in Arisaema Cum Bile were characterized by matching with the local database and comparing with the reference substances and literature information. TCMIP v2.0 was used to obtain the targets corresponding to the identified components of Arisaema Cum Bile and stroke， and the "disease-formula" correlation analysis was carried out to screen the core targets by topological eigenvalues. DAVID 6.8 was used for enrichment analysis of Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway of core targets. According to the "five principles" of Q-markers and combined with literature reports， the Q-markers of Arisaema Cum Bile in the prevention and treatment of stroke were predicted， and the core components acting on these target genes were obtained. Cytoscape 3.8.0 was employed to draw the network diagram of "medicinal materials-active ingredients-target genes-pathways". Finally， AutoDock Vina 1.2.2 was used to calculate and verify the molecular docking between the candidate components and the key targets. ResultA total of 76 chemical components was identified in positive and negative ion modes， 85 core targets were collected for Arisaema Cum Bile in the prevention and treatment of stroke. A total of 31 stroke-related pathways， 23 target genes and 9 main active components of Arisaema Cum Bile acting on these genes were screened， and then we determined 4 possible Q-markers for Arisaema Cum Bile in the prevention and treatment of stroke according to the "five principles". ConclusionThe possible Q-markers of Arisaema Cum Bile for stroke are gallic acid， apigenin-6，8-di-C-glucoside， apigenin and cholic acid， and the target of these four components may be estrogen receptor alpha （ESR1）.