Clinical Metabolomics Study On The Intervention Of Baihe Dihuang Decoction On Yin Deficiency Type Psychological Suboptimal Health State

Rationales: Psychological suboptimal health state(PSHS)is an intermediate state between mental health and mental illness.The incidence rate of PSHS is high and increased year by year,and it may spontaneously into depression,anxiety,sleep disorder and other mental nervous system diseases if without long-term reasonable and timely intervention,which is serious to human health.Therefore,it is of great clinical significance to prevent it from developing into a variety of psychological diseases.There are different syndrome types of PSHS,which is the foundation for the study of diagnostic criteria and drug intervention.Yin deficiency type psychological suboptimal health state is a common classification of PSHS,and the common clinical symptoms is depressed,hung up,abnormal appetite and so on.What' more,its pathogenesis is Yin deficiency and internal heat,which is similar to “Baihe Bing”.Baihe Dihuang decoction(BDD)is a classic prescription treatment of“Baihe Bing”,and it consists of two herbs,namely Lilium pumilum DC.and Rehmannia glutinosa Libosch.Therefore,in the previous study,the self-made diagnostic scale was established,and the yin deficiency type PSHS subjects were selected.The BDD was used to intervene the PSHS subjects,which achieved good clinical results.At the same time,differential metabolites related to the therapeutic effect were selected by using NMR metabonomics.However,the study did not explore the intervention mechanism of BDD.Metabonomics,a branch of system biology,can reflects metabolic changes at the metabolic level.Network pharmacology,which is a combination of multi system biology,multi direction pharmacology and so on,has a strong advantage in the study of multi component multi target multi pathway intervention mechanism of traditional Chinese Medicine.Therefore,the combined of the two methods is helpful to study the active ingredients of BDD,and to find out the related metabolites and their pathway,and to explore its possible mechanism.Objective: To predict the active ingredient and target of BDD;To search and screen the differentially expressed metabolites and their pathways in the clinical plasma samples of PSHS subjects which is treated with BDD;Constructing a “metabolites-metabolic pathway-pathway target-drug targetactive component” network,then associated the activity ingredient and its target predicted targets with the metabolomics results to explore the possible mechanism of BDD in the intervention of yin deficiency type PSHS.Methods: In this study,we used the method of network pharmacology to predict the target of active ingredient of BDD,and screened out the target of the related clinical symptoms in the first.Then,the plasma of PSHS subjects before and after BDD administration were analyzed by metabolomics based on GC-MS and LC-MS.The spectrum of GC-MS and LC-MS were analyzed by XCMS software and Compound Discoverer 2.0,respectively.Noise filtering,peak alignment and extraction were carried out,and finally the retention time,peak area,and m/z ratio data matrix were import into SIMCA-P 14 fore multivariate statistical analysis.Combined the VIP value and SPSS software t-test value,the different metabolites and its pathways related to the effect of BDD were selected.Finally,the results of NMR metabolomics in the previous were integrated with GC-MS and LC-MS results,and the metabolic pathway which was correlated to the intervention effect of BDD was screened.With the method of network pharmacology,the key genes of pathway and the targets of the active ingredient in BDD were integrated.The metabolites,metabolic pathways,targets and active components were combined to study the possible mechanism of BDD on yin deficiency type PSHS.Results: 1.Total 11 active ingredients and their 21 targets were screened in Baihe Dihuang Tang.What's more,the 21 targets mainly involved in gamma-aminobutyric acid signaling pathway,cAMP metabolic process and monoamine transport relevant biological processes.Thus,Baihe Dihuang Tang may play a role of intervention of psychological sub-health by regulating the activity of G-protein coupled amine receptor and the expression of monoamine neurotransmitter.2.The GC-MS metabolomics analysis was used to analyze the plasma of the healthy group and the yin deficiency type PSHS group before and after the intervention.The plasma profile obtained by GC-MS metabolomics were identified,amino acids,organic acids and other 44 endogenous metabolites were identified,and 8 differential metabolites related to the intervention of BDD were screened through a variety of statistical methods of analysis.The analysis of the pathway indicated that the improvement of the clinical symptoms of yin deficiency type PSHS may be mediated by regulating lipid metabolism,energy metabolism and amino acid metabolism.3.On the basis of the previous work,the human plasma LC-MS metabonomics method was optimized,and the plasma of the healthy group and the yin deficiency type PSHS group before and after the intervention were analyzed.14 different metabolites related to the effect of Baihe Dihuang Decoction were found.The related pathway of different metabolites were analyzed,and the result showed that amino acid metabolism and fatty acid metabolism may play an important role of the intervention of BDD.4.The 27 differential metabolites identified by the methods of metabolomics were analyzed,and the metabolic pathways of the 9 important ones were screened out.With the method of network pharmacology,the relationship between the key genes of pathway and the targets of the active ingredient in BDD were analyzed.Finally,the three pathways with the direct interaction between the key genes of pathway and the targets of the active ingredient in BDD were screened.The target GLO1(glyoxalase I),which is the key gene in the pyruvate metabolism pathway of lactic acid,is a key gene of catalpol,rehmaionoside A.MAOA and MAOB,functional target of Stigmasterol and sitosterol,steroidal saponins Compound 10,is the key gene for glutamate metabolism and tyrosine metabolism in metabolic pathways.Among them,phenylalanine is involved in phenylalanine metabolism pathway,3-(3,4-Dihydroxyphenyl)lactic acid is involved in tyrosine metabolism pathway,and tyrosine is involved in two pathways.Conclusion: Baihe Dihuang Decoction mainly involved in amino acid metabolism,glucose metabolism,energy metabolism,fatty acid metabolism and neurotransmitter synthesis and other metabolic pathways.The main pharmacological active components of BDD in the treatment of yin deficiency type PSHS.BDD may regulate the expression of its target GLO1,MAOA,MAOB to improve the symptoms of PSHS,such as depression,anxiety,insomnia,memory loss and so on.The internal integration and external integration analysis of metabonomics is of great significance for the systematic analysis of the mechanism of drug intervention.