| Objective: To develope a research method for the Q-Markers of Euodia rutaecarpa(ER),and confirm its efficacy and/or toxicity Q-Markers.To establish an analytical method for the Q-Markers by high-performance liquid chromatography(HPLC),and provide a basis for quality control of Euodia rutaecarpa.Methods: 1.Full-component analysis of ER was performed by ultra-high performance liquid-quadrupole-flight mass spectrometry(UPLC-Q-TOF/MS).The identification was based on the information such as chromatographic retention,accurate molecular weight,and characteristic fragment ions of the compounds.2.Traditional Chinese medicine serum medicinal chemistry method was introduced to determine the blood components of Euodia rutaecarpa.Combined with the results of the full-component analysis and spectral-effect relationship research in the early stage of the laboratory,the efficacy and toxic components of Euodia rutaecarpa were initially screened out,then the material basis was clarified,finally the efficacy/toxic Q-Markers of Euodia rutaecarpa was preliminarily framed.3.Network pharmacology methods were introduced in this work.With the preliminary determined Q-Marker as the research object,the target prediction and GO enrichment techniques were used to explore the key targets of analgesia,vomiting and toxicity.The main pathways and biological processes were investigated,and the network relationship of component-target-pathway was explored.The mechanism of analgesia,vomiting and toxicity was clarified,and the Q-Markers with clear mechanism was further determined.4.Using the method of HPLC chromatographic fingerprinting and multi-index content determination,the measurability evaluation of the initially determined Euodia rutaecarpa Q-Markers was carried out,and the HPLC method of the main Q-Markers was established to provide a basis for the subsequent quality control research.Results: 1.This article identified 80 chemical constituents from the extract of Euodia rutaecarpa,including alkaloids,limonoids,flavonoids and organic acids.2.In this paper,24 blood components of Euodia rutaecarpa were screened,and the results of the spectral effect study were used to determine the composition of Euodia rutaecarpa Q-Markers.The analgesic components were Evodiamine,Ruteacarpine,Limonin,Dehydroevodiamine,Chlorogenic acid,Hyperoside,Isorhamnetin-3-O-rutinoside,Evodiamine,1-methyl-2-undecyl-4(1H)-quinolone,Trans-caffeoylgluconate.The vomiting ingredients are Limonin,Evodiamine,Hyperoside,Isorhamnetin-3-O-rutinoside,1-methyl-2-undecyl-4(1H)-quinolone.The toxic component is 1-methyl-2-[(6Z,9Z)-pentadienyl]-4(1H)-quinolone,1-methyl-2-indolyl-4(1H)-quinolone,1-methyl-2-[(Z)-6-undecenyl]-4(1H)-quinolo ne,1-methyl-2-[(6Z,9Z,12E)-pentadecalenyl ]-4(1H)-quinolone,Trans-caffeoyl gluconic acid,Ruteacarpine and Evocarpine.3.The 10 analgesic components of Euodia rutaecarpa may regulate vascular endothelial growth factor,protein kinase,and Toll-like cells through 107 analgesic-related targets such as BACE1,DPP4,IL2,MAPK14,MMP12,PIM1,INSR,KIT,MET,and EGFR.Receptors and other signaling pathways,which participate in the process of oxidative stress and inflammatory response to play a role in the treatment of pain.Euodia rutaecarpa ’ s four anti-vomiting components regulate hypoxia-inducible factors and phosphodiesters through 29 vomiting-related targets such as MMP3,CASP3,GSTP1,PDE3 B,MMP9,DPP4,ABO,EGFR,KIT,PDE4 B,ESR1,THRA,and IGF1.Enzymes and other signaling pathways,which are involved in the positive regulation of RNA polymerase II promoter transcription,the positive regulation of tyrosine phosphorylation of Stat5 protein,and the active regulation of nitric oxide biosynthesis.The eight toxic components of Euodia rutaecarpa may be involved in the regulation of Ras protein,Fox O and other signaling pathways through 69 toxic related targets such as CASP1,CASP3,CASP7,SRC,STAT1,CDK6,ANXA5,DPP4,GSTA1,MAPK14.And biological processes such as cell proliferation produce toxicity.4.The methodological investigation showed that: Chlorogenic acid,Hyperoside,Isorhamnetin-3-O-rutinoside,Dehydroevodiamine,Evodiamine,Ruteacarpine,Evocarpine,Dihydroevocarpine in 70.5 ~ 1128 μg / m L,16.2 ~ 259.2 μg / m L,11.6 ~ 184.7 μg / m L,93.6 ~ 1500.8 μg / m L,20.4 ~ 326.4 μg / m L,20.23 ~ 363.45 μg/m L,23.2~371.2 μg/m L,7.07~113.13 μg/m L showed a good linear relationship with the peak area integral values;the sample recovery rates were 101.9 %,97.7 %,99.4 %,99.3 %,101.2 %,98.7 %,100.8 %,101.1 %,RSD values are 2.1 %,2.4 %,2.6 %,2.7 %,2.1 %,2.2 %,1.5 %,2.7 %.Conclusions: In this paper,the “componet-effect-moving” is firstly discussed to explore the pharmacodynamic basis of Euodia rutaecarpa.The mechanism of active ingredients was studied by network pharmacology.Finally,the HPLC measurability evaluation of Euodia rutaecarpa effect Q-Markers was carried out,and Euodia rutaecarpa was identified.The efficacy/toxicity of Q-markers and the establishment of HPLC methods for the relevant Q-Markers.The Q-Markers research method established in this paper reflects the overall and comprehensive characteristics of traditional Chinese medicine,and can provide ideas and methods for the quality evaluation of traditional Chinese medicine.The determination of Euodia rutaecarpa’s efficacy/toxicity Q-Markers can provide a basis for Euodia rutaecarpa’s follow-up research and development. |
