The Contribution Of Radix Bupleuri-Radix Paeoniae Alba To Xiaoyaosan Prescription On Antidepressant Based On Metabolomics

Ratinales: Depression,a global disease with various pathogenic factors,is seriously affecting human physical and mental health.In Chinese traditional medical theory,the disease belongs to the category of "Yu Zheng".At present,development of new antidepressant drugs based on traditional Chinese medicine prescriptions has become a research hotspot due to the increasingly prominent adverse reactions of antidepressant chemicals.However,due to the complexity of traditional Chinese prescriptions.In recent years,our research group has studied the material basis of Xiaoyaosan's antidepressant effect by chemical composition attribution,pharmacokinetics and serum pharmacochemistry methods.It was found that the monarch medicine Radix Bupleuri and the minister medicine Radix Paeoniae Alba contributed a lot to Xiaoyaosan.According to traditional Chinese medicine theory,Radix Bupleuri,Xinsan,Shuganjieyu,and Radix Paeoniae Alba,Suanshou,Yangxuerougan,they are basic herbs of many famous shuganjieyu prescriptions such as Chaihu-shugan-san form �Jingyue's Complete Works�,Sinisan form �Treatise on Cold Pathogenic Diseases�,Xiaoyaosan form �Prescriptions of the Bureau of Taiping People's Welfare Pharmacy�,they are the main drugs in the prescriptions.Researchers analyzed the medication regularity of patent prescriptions of traditional Chinese medicine for depression,the results showed that the frequency of Radix Bupleuri was the first and Radix Paeoniae Alba was the third,The most commonly used herb-pair was Radix Bupleuri-Radix Paeoniae Alba herb pair.Applying LCMS/MS technology and background subtraction method,our team has completed the chemical mechanism study of Radix Bupleuri-Radix Paeoniae Alba herb-pair on the core position in Xiaoyaosan.At the same time,the researchers found that Radix Bupleuri-Radix Paeoniae Alba herb-pair has a clear antidepressant effect when used as a whole.But what is the core position of Radix Bupleuri-Radix Paeoniae Alba herb-pair in the Shuganjieyu prescription? What pharmacological mechanism does Radix Bupleuri-Radix Paeoniae Alba herb-pair exert antidepressant effects? The above questions are not clear.In response to the above problems.In this study,the behavioral wad used to compare the anti-depressant effects of Radix Bupleuri-Radix Paeoniae Alba herb-pair,Xiaoyaosan and Negative group(Xiaoyaosan without Radix Bupleuri and Radix Paeoniae Alba)on CUMS depression rats.The similarities and differences of drugs in the regulation of depression-related biological standards using metabolomics analysis.Metabolic pathway analysis of drug regulation revealed the contribution of Radix Bupleuri-Radix Paeoniae Alba to Xiaoyaosan.From the pharmacological,the core position of herb pair in the compound is revealed.At the same time,based on metabolomics technology,the antidepressant mechanism of Radix Bupleuri-Radix Paeoniae Alba was clarified.Objective: 1.The contribution of Radix Bupleuri-Radix Paeoniae Alba herb pair to Xiaoyaosan was evaluated by comparing the anti-depressant effects of Radix Bupleuri-Radix Paeoniae Alba,Xiaoyaosan and negative group on the behavior(body weight,open field test,sucrose preference rate and immobility time in forced swimming test)on CUMS model rats.2.The serum and liver metabolic profiles of control group,model group,Radix Bupleuri-Radix Paeoniae Alba group,Xiaoyaosan group and negative drug group were studied by combining 1H-NMR and LC-MS/MS metabonomics techniques.The number and level of regulation of three drugs on depression-related biomarkers were compared and analyzed.The contribution of Radix Bupleuri-Radix Paeoniae Alba herb pair to Xiaoyaosan was elucidated by the metabolomics.3.Based on the analysis of metabolites,the metabolic pathway of the biomarkers related to depression,the markers regulated by Radix Bupleuri-Radix Paeoniae Alba,Xiaoyaosan and negative group were analyzed,and the contribution of Radix Bupleuri-Radix Paeoniae Alba herb-pair to Xiaoyaosan was revealed from the level of metabolic pathway.Content and methods: 1.Pharmacodynamic analysis: The experimental rats were randomly divided into control group,model group,positive drug group(venlafaxine hydrochloride 35 mg.kg-1),Radix Bupleuri-Radix Paeoniae Alba herb-pair high and low-doses group(herb amount 15,7.5 g.kg-1),Xiaoyaosan high and low-doses group(herb amount 46,23 g.kg-1),negative control high and low-doses group(herb amount 31,15.5 g.kg-1).Rats were administered at 10 ml�kg-1for each group.The success of the model was verified by comparing the body weight,open field,sucrose preference and forced swimming test of rats in control and model groups,and the effects of Radix Bupleuri-Radix Paeoniae Alba,Xiaoyaosan and negative groups were compared by these behavioral indicator.2.Metabolomics analysis: The serum and liver of rats were analyzed by1H-NMR and LC-MS/MS.Principal component analysis(PCA),orthogonal signal correction filtered partial least squares discrimination analysis(OPLS-DA)and "V-S-plot" were used to screen depression-related biomarkers.The numbers and level of biomarkers regulated by Radix Bupleuri-adix Paeoniae Alba herb-pair,Xiaoyaosan and Negative group were analyzed.3.Metabolic pathway analysis: On the basis of depression-related biomarkers,the similarities and differences of the metabolic pathways regulated by Radix Bupleuri-adix Paeoniae Alba,Xiaoyaosan and Negative group were compared.Results: 1.In the body weight test,weight of the rats was significantly increased in drug-administered groups compared with model group,and there were no significant difference in Radix Bupleuri-adix Paeoniae Alba,Xiaoyaosan and negative group.In the open field test,All drug-administered groups could regulate the spatial activity of rats.Among them,The high-dose group Xiaoyaosan has a slightly stronger effect on the number of crossing than the Radix Bupleuri-adix Paeoniae Alba,the low-dose group was equivalent and there was no significant difference between the groups.There was a significant difference between negative group and the Xiaoyaosan group in the low-dose group.In the sucrose preference test,The high-dose group Radix Bupleuri-adix Paeoniae Alba was equivalent to Xiaoyaosan,and the low-dose group had a slightly stronger callback,and there was no significant difference between the groups.In the forced swimming experiment,the rats' immobility time of Radix Bupleuri-adix Paeoniae Alba-treated was shorter than Xiaoyaosan and the negative group.2.Serum metabolomics results: In the serum metabolic profile analysis based on 1H-NMR and LC-MS/MS techniques,the blank group and the model group were significantly separated in the PCAand OPLS-DAmaps,indicating that the serum metabolic profiles of the model group rats were significantly changed compared with the control group.Xiaoyaosan and the herb-pair could ameliorate 22 and 17 metabolites respectively,and the negative control could only ameliorate 13 metabolites.3.The metabolic pathway of serum includes: Pathway analysis of biomarkers depression-related revealed a total of 15 metabolic pathways(influence value(29)0.1 was considered as a depression-related metabolic pathway).In terms of the number of total metabolic pathways,the number of metabolic pathways regulated by Radix Bupleuri-adix Paeoniae Alba,the Xiaoyaosan and the negative group was equivalent;The important metabolic pathways regulated by the three drugs,the importance and significance of the metabolic pathways regulated by Radix Bupleuri-adix Paeoniae Alba and Xiaoyaosan aremore similar.4.Liver metabolomics results: In the serum metabolic profile analysis based on 1H-NMR and LC-MS/MS techniques,the control group and the model group were significantly separated in the PCAand OPLS-DAmaps,indicating that the serum metabolic profiles of the model group rats were significantly changed compared with the control group.Xiaoyaosan and the herb-pair can ameliorate 25 and 24 metabolites respectively,and the negative control can only ameliorate 14 metabolites.5.The metabolic pathway of liver includes: Pathway analysis of depression-related biomarkers revealed a total of 10 metabolic pathways.In terms of the number of total metabolic pathways,9 of them were significantly regulated by Radix Bupleuri-Radix Paeoniae Alba and Xiaoyaosan,and 6 of them were significantly regulated by the negative group.The importance and significance of Radix Bupleuri-adix Paeoniae Alba and Xiaoyaosan regulating metabolic pathways are moresimilar.Conclusin: This study reveals the contribution of Radix Bupleuri-Radix Paeoniae Alba to Xiaoyaosan from three levels: behavioral,metabolite and metabolic pathways.At the behavioral level,Radix Bupleuri-adix Paeoniae Alba showed an antidepressant effect similar to Xiaoyaosan on most indicators;At the metabolite level,Radix Bupleuri-adix Paeoniae Alba herb-pair was closer to Xiaoyaosan in the number and level of markers;At the metabolite pathway level,the number of Radix Bupleuri-Radix Paeoniae Alba herb-pair in the regulation of total metabolic pathways and important metabolic pathways were more similar to Xiaoyaosan.The importance and significance of metabolic pathways regulated by herb-pair and Xiaoyaosan wree more similar.