Investigation On The Metabolism Process Of The Effective Constituents Of Sini Powder With Antidepressant Effect In Vivo Based On Processing And Compatibility

In this paper,based on the research idea of "individual herb,herb-pair,herbal formula",taking the representative drugs Bupleuri Radix(BR)and Paeoniae Radix Alba(PRA)as examples,Sini Powder(SNP)as the whole prescription,and depression as the disease,the metabolism process of antidepressant active components in vivo before and after vinegar-processing of BR and PRA and compatibility prescription of Sini Powder was investigated.A rapid and sensitive ultra-high performance liquid chromatography-mass spectrometry(UHPLC-QqQ-MS/MS)method was successfully applied to a holistic comparative pharmacokinetic investigation of antidepressant active constituents in SNP based on three different forms of drug composition(individual herb,herb-pair,herbal formula)from BR and PRA before and after vinegar-processing in the plasmas of depressive rats.According to the results of dynamic changes in vivo obtained above,the effects of intragastric administration groups on the mRNA expression of three CYP450 subtype enzymes in the livers of depressive rats was further studied.The qualitative analyses of metabolomics in depressive rats of intragastric administration groups were carried out to explain the absorption,distribution,metabolism,and excretion process of the drugs entering the body due to the intervention system being affected by organisms.Finally,the endogenous metabolomics of the intervention depressive rats in intragastric administration groups was studied from the perspective of endogenous biological targets,and the overall metabolic pathways of SNP against depression was confirmed from the response regulation of the biological organism caused by the intervention system,and the potential biomarkers related to the onset of depression were screened and identified.According to the changes of biomarkers after modeling and administration,the mechanism of regulating imbalance was discussed.In chapter one,fourteen constituents,such as saikosaponin a(SSa),saikosaponin b2(SSb2),saikosaponin c(SSc),saikosaponin d(SSd),paeoniflorin(Pae),albiflorin(Alb),benzoylpaeoniflorin(Ben),oxypaeoniflorin(Oxy),hesperidin(Hsd),hesperetin(Hst),naringin(NG),liquiritin(LN),glycyrrhizic acid(GA),and liquiritigenin(LG),were selected as detection indicator.The study was based on comparative pharmacokinetic investigation of antidepressant active ingredients in SNP in plasmas of chronic unpredictable mild stress(CUMS)depressive rats based on processing and compatibility.The depressive model was established by CUMS for 4 consecutive weeks,and the behavioral indices were investigated by sugar water preference experiment,open field experiment,and forced swimming experiment.The rats that were successfully modeled were randomly divided into BR group,vinegar-processed BR(VPBR)group,PRA group,vinegar-processed PRA(VPPRA)group,herb-pair(BR-PRA)group,herb-pair(VPBR-VPPRA)group,herbal formula(SNP)group and herbal formula(SNP containing VPBR and VPPRA)group for pharmacokinetic analysis.The results showed that there were significant differences in the Cmax,AUCo-24,AUC0-∞,t1/2,and MRT0-24 of some constituents(saikosaponins and total glucosides of paeony)among the three different forms of drug composition from BR and PRA before and after vinegar-processing.At the same time,the AUC0-24,AUC0-∞,and Cmax of LN,GA,LG,Hsd,and Hst in the group of SNP containing VPBR and VPPRA were significantly increased than those in the SNP group(P<0.05,P<0.01).;In chapter two,PCR technology was employed to further explore the effects of intragastric administration groups on the mRNA expression of three CYP450 subtypes(CYP1A2,CYP2E1,and CYP3A1)in depressive rats.The experimental results showed that depression caused by CUMS had the strong inducing effects on CYP1A2,CYP2E1,and CYP3A1.The mRNA expression of CYP1A2,CYP2E1,and CYP3A1 in the livers of rats under depression could be recovered to a certain extent after vinegar-processing and compatibility with BR and PRA.In addition,the regulation effects of herb-pair groups on the mRNA expression of CYP1A2,CYP2E1,CYP3A1 were more obvious than those of individual herb groups,and the regulation effects of herbal formula groups were more obvious than those of individual herb and herb-pair groups,indicating that "BR-PRA" herb-pair played an important role in SNP.Among them,it was also found that the principal and assistant drugs(BR and PRA)in SNP had different contributions to the regulation of the mRNA transcription levels of drug metabolism enzymes,which also proved the rationality of BR as principal drug in SNP.Based on the experiments of chapter one and chapter two,chapter three was innovatively focused on SNP with multi-constituents and multi-targets,and the split prescriptions from three levels of "individual herb-herb-pair-herbal formula " were investigated,and the two levels of"crude and vinegar-processed products" were compared.Based on UPLC-Q-TOF/MS technology,the differences of metabolites in plasmas,biles,urines,and feces of fourteen antidepressant constituents before and after vinegar-processing of BR and PRA and compatibility prescription of Sini Powder were investigated.Firstly,the distributions of the prototypes of these fourteen constituents in depressive rats of each administration group were identified.And 27 metabolites from BR,30 metabolites from PRA,52 metabolites from BR-PRA,118 metabolites from SNP,and 168 metabolites from SNP containing VPBR and VPPRA were identified as well.Secondly,it was speculated that these constituents were mainly existed in the forms of prototype,deglycosylation,deoxygenation,demethylation,oxidation,methylation,glucuronidation,sulfation,and others in depressive rats of each administration group.In depressive rats,there were differences in metabolites between the "individual herb-herb-pair-herbal formula" before and after compatibility and the "herbal formula-herbal formula containing processed products"before and after processing.In chapter four,the metabolomics technology of UHPLC-Q-TOF/MS was used to explore the pathogenesis of depression,and the differences in plasma metabolic profiles of depressive rats after different drug intervention were evaluated.Using stoichiometry and the establishment of a discriminant model,sixteen kinds of differential metabolites related to depression were screened out.The appearance of these differential metabolites indicated that there were abnormalities in amino acid metabolism,lipid metabolism,energy metabolism,and nucleotide metabolism in depressive rats,and SNP may intervene depression by regulating lipid metabolism and nucleotide metabolism.The interventions of herb-pair and individual herb were achieved through energy metabolism,nucleotide metabolism,and lipid metabolism,respectively,while the interventions of SNP containing VPBR and VPPRA were achieved through amino acid metabolism,lipid metabolism,energy metabolism,and nucleotide metabolism.