To Study The Compatibility Mechanism Of Coptidis And Magnolia Officinalis Based On Composition And Pharmacodynamics

Basis of selection:The study of the compatibility mechanism of traditional Chinese medicine is a key issue in the modernization of traditional Chinese medicine.However,due to the complexity of traditional Chinese medicine components,the study of traditional Chinese medicine compatibility faces many obstacles.At present,the study of TCM compatibility is generally carried out from three parts: composition,pharmacodynamics and pharmacokinetics,and is carried out at a single level,which cannot fully explain the compatibility mechanism.The compatibility of Coptis coptidis and magnolia officinalis was derived from "Houpu Wan" in "Taiping Shenghui Prescription".Coptis coptidis bitter cold and magnolia officinalis xinwarming are used to treat diarrhea.As the material basis of Chinese medicine,the composition is the breakthrough point of modern research on the compatibility of Coptis and Magnolia officinalis.Coptis coptidis and magnolia officinalis are combined to treat diarrhea,and modern studies have proved that this medicine can treat inflammatory bowel disease.Purpose:Based on the in vivo and in vivo chemical composition changes and in vivo action process before and after the compatibility of Coptidis and Magnolia officinalis,combined with the overall study of the mechanism of the compatibility of pharmacodynamics,traditional Chinese medicine and modern pharmacology combined to provide theoretical basis for the clinical use and further research and development of Chinese medicine pairs.It also provides exploratory ideas for the study of other drug pairs and TCM compounds.Method:(1)UPLC-Q-Exactive-Orbitrap-MS technique was used to study the changes of chemical constituents in vitro in single and co-decoction of Coptis coptidis andMagnolia officinalis,providing directions for further research on the pair of Coptis coptidis and Magnolia officinalis.(2)The mouse serum samples were analyzed and identified by UPLC-Q-Exactive-Orbitrap-MS technique to study the changes of blood components before and after drug combination;Combined with multivariate statistical analysis,the different metabolites and related metabolic pathways of Radix coptidis and Magnolia officinalis were obtained.(3)Colitis mouse model was established and divided into blank group,model group,positive control group,coptis magnolia officinalis low-dose,medium-dose and high-dose groups,coptis officinalis group and magnolia officinalis group.After 14 days of continuous administration,serum and colon tissues were collected and pharmacodynamic results of each group were determined.(4)colonic tissues of mice were stained with H&E,and morphological changes of each group were observed.The expression of TLR4 protein was detected by immunohistochemical section.The concentrations of TNF-α,IL-1β,IL-6 and IL-10 in serum were determined by ELISA.The contents of TLR4,IL-6,TNF-α and IL-1βin colon tissues of mice were detected by RT-PCR.The expressions of TLR4,NF-κβ and MYD88 in colon tissues were detected by Western Blot.Results:(1)UPLC-Q-Exactive-Orbitrap-MS technology was used to detect the classification and change of chemical constituents in the single and co-decoction of Coptis officinalis and Magnolia officinalis.Most of Coptis officinalis were alkaloids,and the peaks were higher in the positive ion mode,while most of magnolia officinalis were phenylpropanes,and the peaks were higher in the negative ion mode.With the appearance and disappearance of chemical components,the content of index components in the drug pair was lower than that in the single drug.(2)Based on UPLC-Q-Exactive-Orbitrap-MS technique,27 prototype components were identified,including 12 components of coptis berberine,palmatine, pharmacoglycorine,and 11 components of magnolia officinalis,honokiol,and honokiol.Coptis coptidis separately regulated 23 differential metabolites,Magnolia officinalis separately regulated 14 differential metabolites,and 57 metabolites after compatibility.In terms of quantity,the drug pair was more than the single drug.The results of metabolic pathway showed that the drug pair may exert pharmacological effects through influencing tricarboxylic acid cycle and amino acid metabolism.(3)After the intervention of Coptidis and Magnolia officinalis,the physical signs of IBD mice gradually recovered;HE staining showed that Coptis coptidis,magnolia officinalis and drug pair could improve colon tissue injury and alleviate pathological changes.ELISA results showed that Coptis officinalis,Magnolia officinalis and their drug pairs could significantly inhibit TNF-α,IL-1β,IL-6 and enhance IL-10 levels(P < 0.05).Immunohistochemical results showed that TLR4 protein expression could be inhibited in each administration group(P <0.05).RT-PCR results showed that compared with model group,the expression multiples of TLR4,IL-6,TNF-α and IL-1β in drug administration groups were significantly decreased(P < 0.05).The results of WB showed that the expressions of TLR4,MYD88 and NF-κB in all administration groups were down-regulated(P< 0.05),and the high dose group had the best effect,followed by the medium and low dose groups,and the effect of the same dose group was stronger than that of single drug.Conclusion:(1)The pharmacodynamic material basis of Coptidis-Magnolia officinalis pair was clarified through in vitro chemical composition detection and verification before and after the compatibility of Coptidis and magnolia officinalis.It was found that the pharmacodynamic material basis was related to the content of Coptidismagnolia officinalis and the disappearance and formation of chemical components,which provided certain basis for further study of the in vivo action process and pharmacodynamic process of Coptidis-magnolia officinalis.(2)Through the detection and analysis of chemical components in vivo before and after Coptidis and Magnolia officinalis incompatibility,it is speculated that Coptidis,magnolia officinalis and drug pairs may play their effects based on absorption and metabolism of chemical components,regulation of differential metabolites and metabolic pathways,which lays the foundation for further pharmacodynamic studies.(3)Pharmacodynamics showed that Coptidis magnolia officinalis inhibited inflammatory response by regulating TLR4/MYD88/NF-κB pathway,and the combined efficacy of Coptidis magnolia officinalis was higher than that of any single drug,and the efficacy of the drug pair group was dose-dependent.