| Traditional Chinese medicine has been used in my country for thousands of years.Its components are complex,and its targets are many.It is often the coexistence of efficacy and toxicity.Through practice,countless sages of traditional Chinese medicine have formed unique traditional Chinese medicine theories such as processing and compatibility to achieve reconciling medicine,reducing toxicity and increasing efficiency.Euodiae Fructus(EF).,pungent,bitter,hot,mildly toxic,its fresh smell is strong and irritating,and it is often used as medicine after aging in clinical practice.The sayings about the aging of EF as medicine are in records in many traditional Chinese medicine classics,but there is no scientific explanation for the ageing of EF.This project is based on the theory of “the older,the better” of EF,and conducts a comprehensive study on the composition of EF before and after aging,combined with chemometric analysis methods to establish a quality discrimination method,and builds a“component-target-pathway” network with the help of network pharmacology,To scientifically explain the theory of “the older,the better” from the perspective of material basis-mechanism of action.(1)EF is a variety of original medicinal materials(Euodia rutaecarpa(Juss.)Benth.,Evodia rutaecarpa(Juss.)Benth.var.officinalis(Dode)Huang.or Evodia rutaecarpa(Juss.)Benth.var.bodinieri(Dode)Huang.),this experiment will study its origin.First,establish a Heracles Ⅱ ultra-fast gas phase electronic nose method to detect the odor of EF,detect and characterize its volatile components.Then,an unsupervised pattern recognition model(PCA)and a supervised pattern recognition model(PLS-DA)were established,and the index components that affected their classification were determined,and the built model was verified through a validation set.The experimental results show that myrcene,α-pinene,andβ-elemene can be used as reference indicators to distinguish the three types of EF.(2)From the perspective of the non-volatile components of EF,the material basis of EF“the older,the better” theory was explored.Collected 22 batches of Evodia in different years to form different batches of aging groups,and 6 batches of Evodia in the same year constituted the same batch of aging groups.The two groups of samples were studied in parallel.First,establish the HPLC fingerprint of Evodia and analyze the fingerprint information of Evodia by chemometrics.The results show that different batches of aging group:specifications have a significant impact on the quality of Evodia,22 batches of different batches of Evodia have a clear boundary according to specifications.The content of alkaloids and phenolic acids in the three specifications of EF is quite different.Eliminating the influence of specifications,the PCA analysis showed that the samples were grouped into long-term storage and short-term storage.The same batch of aging group:Evodia edulis aging for 0-15 months has no obvious influence on its fingerprint information,and the origin factor has a significant influence on the quality of medicinal materials.Then,in order to further determine the changes in the composition of the aging process,the research was carried out from two levels of the alcohol extract and water extract of EF.The results showed that the content of evodiamine and limonin in the ethanol extract of EF decreased with the aging time(P<0.05);the content of neochlorogenic acid,chlorogenic acid,and hyperoside in the water extract of EF all decreased(P<0.05),the content of caffeic acid increased(P<0.05).(3)From the perspective of the volatile components of EF,the material basis of EF “the older,the better” theory was explored.With the help of Heracles Ⅱ ultra-fast gas phase electronic nose technology,the gas phase fingerprints of EF are obtained and combined with chemometric models(PCA,DFA,SQC,SIMCA),the volatile components of EF(same batch,different batches)are analyzed for different aging time.Experiments have found that in different batches of aging group: the aging time has a significant effect on its odor,and the chemometric model can accurately identify the long-term and short-term aging of EF.The same batch of aging group: the odor components of EF are related to the ageing time,and the chemometric model can better identify the aged and unaged EF.(4)On the basis of Experiment 2 and Experiment 3,the different components of EF after aging were obtained,and the molecular mechanism of EF “the older,the better” was studied with the help of network pharmacology.First,use the TCMSP database and the Swiss Target Prediction database to obtain the relevant gene targets of EF before and after aging,and then construct the protein interaction through the protein interaction database(STRING),and screen the core target proteins in the Cytoscape software to construct a protein interaction network Visualize the network,and finally,use the David database for GO analysis and KEGG signal pathway enrichment analysis.In this experiment,based on 11 different components,a total of 75 potential targets,268 gene function annotations and 42 target biological pathways were predicted.The results of network pharmacology experiments found that it is speculated that limonin,β-phellandrene,hyperoside,and dehydroevodialine can affect Hepatitis C,NAFLD,Drug metabolism-cytochrome P450 pathway has a regulatory effect on the attenuation of evodia;dehydroevodialine can regulate the Neuroactive ligand-receptor and Calcium signaling pathway Plays a protective effect on the nervous system and thus has a synergistic effect.Through the research of this subject,a method for quickly identifying the origin of EF by the Heracles Ⅱ ultra-fast gas phase electronic nose was constructed;the analysis method of the non-volatile and volatile components in Evodia was constructed and applied to the chemical composition of the aging process of EF.Research on the law of change;construct the “component-target-pathway” network before and after aging of Evodia edulis and explain its mechanism of action.This research has important reference significance for the research of aging traditional Chinese medicine. |
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