Study On The Pharmacodynamics And Metabolomics Of Zhizihoupo Decoction And Its Single Decoction In The Treatment Of Depressio

Depression is currently one of the major medical challenges,affecting more and more people.Compared with western medicine,traditional Chinese medicine(TCM)has the advantages of gradual effectiveness,less side effects,and higher safety for long-term use in treating depression.Zhi-Zi-Hou-Po decoction(ZZHPD),which was first recorded in Zhang Zhongjing’s "Shanghan Lun",is composed of three herbs:Gardeniae Fructus(GF),Aurantii Fructus Immaturus(AFI),Magnoliae Officinalis Cortex(MOC)and is mainly used to treat depression such as chest and abdominal fullness caused by heat accumulation.Modern clinical studies have shown that GF,AFI,MOC and ZZHPD have certain therapeutic effects in treating depression.Although there has been some progress in the research on ZZHPD,the material basis and mechanism of action of single herbs and combined decoction(co-decoction)are still unclear.Therefore,in-depth research on the mechanism of action of the co-decoction and single decoction of ZZHPD in treating depression based on the theory of co-decoction,can help to comprehensively understand the pharmacological effects and efficacy of TCM,and further expand its clinical application and development.Based on literature review and previous work,this project utilizes the theory of codecoction,a method of TCM preparation where multiple herbs are simultaneously boiled in the same pot of water,to fully exploit the therapeutic effects of TCM,strengthen the synergy between herbs,and enhance the efficacy of TCM formula.The potential key pharmacological components of the co-decoction and single decoction of ZZHPD will be preliminarily screened using liquid chromatography-mass spectrometry(LC-MS)technology combined with serum pharmacokinetic theory.On the other hand,the mechanism of action of ZZHPD and single decoction will be analyzed through metabolomics,exploring the endogenous differential metabolites in depressed mice and the metabolic changes after drug intervention,and constructing a metabolic regulatory pathway for the antidepressant effects of ZZHPD and single decoction.This will help to clarify the mechanism of action of ZZHPD and single decoction at the metabolic level,gain a deeper understanding of their pharmacological effects and drug interactions from the perspective of co-decoction,and guide the rational use and clinical treatment of TCM.In order to clarify the pharmacological effects of ZZHPD and its single decoction,this study used UPLC-QE-Orbitrap-MS technology to separate the liquid of the single medicine in ZZHPD,the co-decoction of the two medicines and the three medicines.The co-decoction of different medicines was analyzed and compared,and the subsequent data processing was carried out to excavate the chemical components in the co-decoction and single decoction of ZZHPD,as well as the newly formed unknown compounds.Further investigation of ZZHPD and single decoction of blood migration components and components into the brain,the results found that 30 components into the blood and 20 components into the brain.Among the components that entered the brain,5 ingredients were traced to GF,11 ingredients were traced to AFI,5 ingredients were traced to MOC,and 1 ingredient was only found in ZZHPD.The chronic unpredictable mild stress(CUMS)combined with single housing was used to establish an ICR mice model of depression.The mice were divided into six groups:a control group,a CUMS model group,a group treated with a combined decoction of ZZHPD,a group treated with a single decoction of GF,a group treated with a single decoction of AFI.and a group treated with a single decoction of MOC.The mice were evaluated using sucrose preference test(SPT),open field test(OFT),and novelty suppressed feeding test(NSF test).The results showed that compared to the model group,the mice in the control group and the groups treated with GF,AFI,MOC,and the combined decoction of ZZHPD exhibited a significant increase in sucrose preference(P<0.05).a significant decrease in latency to feed(P<0.05),a significant increase in the exploration time of the central area,and a significant increase in total distance traveled(P<0.05).Additionally,the combined decoction of ZZHPD significantly reversed the decrease in exploration time of the central area in the model group mice,while the group treated with AFI significantly increased the total distance traveled by the mice(P<0.05).Hematoxylin-eosin(HE)showed that in the model group mice,there was more separation between the mucosal epithelium and the lamina propria,and a small amount of mucosal epithelium shedding was observed,while the administration of the GF,AFI,and MOC and the co-decoction group of ZZHPD could repair this damage to a certain extent,and the repair effect of the co-decoction of ZZHPD was significantly better than that of the single decoction.The results showed that both the co-decoction and single decoction of ZZHPD had certain improvement effects on the behavioral and intestinal tissue morphology of CUMSinduced depressive mice.In addition,the mechanism of the ZZHPD in treating depression was explored using fecal metabolomics.The overall regulatory mechanism of ZZHPD and its decoction intervention in the CUMS model mice were evaluated through fecal samples.As a result,82 differential metabolites were screened and identified,of which 36 pharmacologically active substances,such as 5-Hydroxyindoleacetate and N-Myristoyl Asparagine,were discovered in the GF group.These substances mainly play a role through 10 pathways,such as the metabolism of valine,leucine,and isoleucine,purine metabolism,and tryptophan metabolism.In the AFI group,20 pharmacologically active substances,such as Dihydrogenistein,Dihydroretrofractamide B,and 5-L-Glutamyl-taurine,were discovered.These substances mainly play a role through 10 pathways,such as the metabolism of valine,leucine,and isoleucine,purine metabolism,and pyrimidine metabolism.In the MOC,4 pharmacologically active substances,such as Dihydroretrofractamide B,L-Ornithine,and L-Isoleucine,were discovered.These substances mainly play a role through 6 pathways,such as the metabolism of valine,leucine,and isoleucine,and arginine biosynthesis.27 pharmacologically active substances,such as Enterodiol,13’Hydroxy-alpha-tocotrienol,and Esculentic acid,were discovered in the ZZHPD.These substances mainly play a role through 8 pathways,such as glutathione metabolism,arginine and proline metabolism,the degradation of valine,leucine,and isoleucine.In summary,this study explores the therapeutic effects of the combined decoction theory and the single decoction method of ZZHPD in treating depression.The research examines the pharmacological substances and mechanisms of action,and ultimately discovers that both ZZHPD and the single decoction method have a certain effect on treating depression.Comprehensive analysis of metabolomics indicates that L-isoleucine is a key biological marker for depression in both the combined decoction and single decoction,and the regulation of valine,leucine,and isoleucine metabolism is an important pathway for the therapeutic effect of ZZHPD on depression.