| Traditional Chinese medicine compound is an important means for clinical treatment of diseases in traditional Chinese medicine.Its clinical application has a history of more than 2,000 years.Even in the era when Western medicine is developing so rapidly,Chinese medicine plays a very important role in the treatment of chronic diseases or disease recovery.Metabolomics aims to study the mechanism of action of drugs by analyzing small molecule compounds in metabolites and backtracking the pathological and metabolic pathways that have occurred.The application of metabolomics technology to explore the mechanism of action of traditional Chinese medicine provides a new idea for the study of Chinese herbal compound and its compatibility.The compound Chinese medicine Jinlingzi San is made up of the same amount of Toosendan Fructus and Corydalis yanhusuo.The first appearance is in Liu Wansu's "Su Wen,Disease,Qi and Gas Insurance",which is the treatment of liver stagnation.A representative formula commonly used in various pain disorders.Currently mainly used for the treatment of gastric and duodenal ulcers,chronic gastritis,chronic hepatitis,cholecystitis and other diseases.Domestic research on Jinlingzisan is mainly based on drug ratio,processing method and pharmacological research.The research on its treatment mechanism is still unclear.In order to explore the mechanism and metabolic pathway of Jinlingzi San and its disassembled in the treatment of diseases,this study established a model of foot swelling caused by acetic acid writhing,hot plate and carrageenan,combined with gas chromatography-mass spectrometry(GC/MS)and liquid chromatography-mass spectrometry(HPLC/MS)techniques were used to find related differential metabolites,construct metabolic network maps,and reveal the anti-inflammatory and analgesic mechanism of Jinlingzi san from the metabolic level.1.Research on the quality standard of Jinlingzi SanIn order to ensure the quality of Jinlingzi San is controllable,the content of the main active ingredients of the two traditional Chinese medicines in Jinlingzi San is determined,which lays a foundation for the pharmacodynamic study of Jinlingzi San for anti-inflammatory and analgesia.In this study,the method for determination of Toosendan Fructus and Corydalis yanhusuo chinensis recorded in the 2015 edition of Chinese Pharmacopoeia was improved by using Toosendan Fructus and Tetrahydropalmatine as standard.The content determination and method validation were carried out.The results showed that the content of active ingredients in Toosendan Fructus and Corydalis yanhusuo was in line with the relevant requirements of the Pharmacopoeia.2.Metabolomics study on the analgesic mechanism of Corydalis yanhusuoIn order to explore the mechanism of action of Corydalis yanhusuo in the treatment of pain,this study used an animal model of acetic acid writhing to observe the number of writhing in the mouse within 15 min after modeling,and calculate the writhing inhibition rate to evaluate its analgesic activity.The mouse serum was analyzed by GC/MS,and the collected data was imported into XCMS to obtain a related data matrix.In order to visualize the data,it was introduced into SIMCA-P or MetPA for PCA,PLS-DA,and OPLS-DA analysis.The results showed that the profile of the biometabolites in the control and model group changed significantly,and a total of 18 differential metabolites such as glycine?proline?leucine and citric acid were screened out.Enrichment of metabolic pathways by MetPA,the effects of Corydalis yanhusuo on the treatment of pain-related pathways are glycerolipid metabolism,glyoxylate and dicarboxylate metabolism,glycine,serine and threonine metabolism,linoleic acid metabolism,Valine,leucine and isoleucine biosynthesis.The analgesic mechanism is mainly related to the secretion of the central nervous system and peripheral pain-causing substances.3.Metabolomics study on the analgesic mechanism of Toosendan FructusIn order to explore the mechanism of the effect of Toosendan Fructus on pain,this experiment used a mouse model of chemical stimulation pain(acetate writhing method)to observe its analgesic effect.Pharmacological results showed that the middle dose group of Toosendan Fructus had a good effect on the inhibition rate of writhing in mice,but the analgesic effect was lower in the high dose group,which may be related to the liver and kidney toxicity of Toosendan Fructus.Higher doses may be the mouse body has an effect.Analysis of mouse serum by GC/MS showed that a total of 13 differential metabolites such as alanine?serine and malic acid were screened.The more relevant metabolic pathways are glycine,serine and threonine metabolism,glycerolipid metabolism,glyoxylate and dicarboxylate metabolism,Aminoacyl-tRNA biosynthesis.The mechanism of action is primarily related to the central nervous system pathway.4.Metabolomics study on the analgesic effect of Jinlingzi San and its single-drugIn order to further compare the similarities and differences of Jinlingzi San and its single-drug analgesic mechanism,this experiment used hot plate method and acetic acid writhing animal model to observe the improvement of pain threshold and writhing inhibition rate in mice,and compare the compound Chinese medicine and Its single-drug analgesic effect.The results showed that the analgesic effect of compound Chinese medicine was better than that of unilateral.The serum of the mice was determined by GC/MS technique,and 14 differential metabolites related to pain were screened out such as glycine?threonine and isoleucine.Metabolic pathway enrichment by MetPA revealed that the metabolic pathways are metabolized by glycine?serine and threonine metabolism,Valine?leucine and isoleucine biosynthesis.The metabolic network map drawn showed that the mechanism of compound analgesia was mainly related to the change of opioid content and the secretion of peripheral pain-causing substances.5.Metabolomics study on the anti-inflammatory effects of Jinlingzi San and its single-drugIn order to explore the effects of Jinlingzi San and its single-drug on metabolite changes in inflammatory rat models,this study used carrageenan-induced rat paw swelling model to observe the swelling of the rats in each group after modeling.Tumor necrosis factor alpha(TNF-?),interleukin-6(IL-6)and interleukin-1?(IL-1?)levels in rat serum.The results showed that the degree of swelling of the model group was significantly higher than that of the control group(p<0.05),and the levels of IL-6,IL-1? and TNF-? were significantly increased(p<0.05).The collected serum was analyzed by GC/MS technique,and the urine was analyzed by HPLC/MS.A total of 28 kinds of differential metabolites related to inflammation such as glutamine?alanine and succinic acid were screened from serum and urine.The relevant differential metabolites are network-enriched to obtain metabolic pathways with high correlation between inflammation,such as ?-alanine metabolism,D-glutamine and D-glutamate metabolism.These metabolic pathways are mapped to network diagrams to explain their anti-inflammatory mechanisms.The anti-inflammatory mechanism of Jinlingzi San is mainly immune regulation,inhibiting leukocyte chemotaxis,migration and activation.The most influential factors are cytokine pathway and nuclear transcription factor-?B pathway. |
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