Research On Efficacy Of Chaihu Shugan Powder By Chromatography-mass Spectrometry Combined With Network Analysis

Chaihu Shugan Powder is a famous prescription for regulating “qi” in traditional chinese medicine,which has the effect of regulating liver,promoting blood circulation and relieving pain.At present,most of the researches on Chaihu Shugan Powder focus on the certain kind of active components and their biological activities,but there is little analysis on the compatibility mechanism,in vivo components / in vitro components and pharmacological effects of this prescription.In this study,the chromatography-mass spectrometry and bioinformatic tools were used to analyze the volatile components of the three drug pair of Chaihu Shugan Powder,‘Radix burpleuri-Radix paeoniae alba ’,‘Pericarpium Citri Reticulatae-Fructus aurantii’and ‘Rhizoma Chuanxiong-Rhizoma Cyperi’,so as to explore the internal mechanism of Chaihu Shugan Powder for ‘regulating liver,,activating blood circulation and relieving pain’.(1)In chapter two,LC-Orbitrap MS was used to identify the protocompounds of‘Radix burpleuri-Radix paeoniae alba ’ in intestinal and serum samples,and using the text mining of biological information related to its role in depression through Pubmed database.The potential mechanism of ‘Radix burpleuri-Radix paeoniae alba ’ against depression was discussed from the biological pathway of ‘gut microbiota-gut-brain axis’.There are seven prototype components were detected in the small intestine of rats,but its were trace in serum.And the results showed that‘Radix burpleuri-Radix paeoniae alba ’ could affect the proportion of intestinal flora(Firmicutes,Bacteroidetes)and its metabolites,the Saikosaponin A and Saikosaponin D significantly up-regulated the expression of Tr KB in hippocampus and the release of BDNF in brain neurons.Meanwhile,the paeoniflorin and paeoniflorin can promote the production of BDNF and regulate the content of c AMP and c GMP in the hippocampus.Its the potential pharmacological basis of Chaihu Shugan Powder.(2)In the third chapter,LC-Orbitrap MS and HS-SPME-GC/MS were combined to detect the compounds of ‘Pericarpium Citri Reticulatae-Fructus aurantii’ in serum and small intestine of rats,and the Pharmmapper database and Autodock Vina were used to obtain the potential targets of components.Then,the network enrichment analysis was carried out to the potential targets,which the pathways and organs/tissues relate to targets were determined.Finally,the corresponding diseases and TCM diseases of the target are associated by text mining,and the relationship between ‘compound-disease’ reported in references were supplemented.A total of 15 protocompounds and 6 metabolites of ‘ Pericarpium Citri Reticulatae-Fructus aurantii’ were detected in serum and small intestine of rats,and 6 volatile components were detected in mouse serum.Then,the 15 protocompounds and 2 metabolites(M1,M2)were studied further.The results showed that 17 compounds were associated with 104 potential targets and enriched in 18 pathways,which were related to diseases of circulatory system,digestive system and respiratory system,and corresponding to TCM syndromes of qi stagnation,chest obstruction and tenesmus.And the network of‘Compound-Potential Target’,‘Target-Disease’,‘Target-Pathway’ and ‘Target-Organ/Tissue’ were constructed.Finally,the ‘Five elements theory’ of TCM was used to supplement the relationship between the disease and TCM disease.This chapter describes the compatibility mechanism of ‘Pericarpium Citri Reticulatae-Fructus aurantii’ drug pair on qi stagnation,chest obstruction and tenesmus syndromes,reflected the mechanism of ‘multi-component,multi-target,multi-pathway’.Its the potential pharmacological basis of Chaihu Shugan Powder.(3)In Chapter four,LC-QTOF-MS and HS-SPME-GC/MS were used to identify the compounds in mouse serum,the pharmacophore model prediction of Pharmmapper platform and molecular docking Autodock Vina tool were used to determine the potential targets of compounds in vivo of ‘Rhizoma Chuanxiong-Rhizoma Cyperi’.Then,the corresponding diseases and the related TCM syndromes of potential targets were matched through TTD,TCMSP and Pub Med databases.A total of 38 protocompounds were detected in serum of mice,corresponding to 70 potential targets,which were mainly related to diseases of circulatory system,endocrine system and nervous system,corresponding to qi stagnation and blood stasis syndrome in TCM.Among them,part of the relationship between ‘Compound-Target-Disease’ has been confirmed by reports,and the network topology method was used to analyze the ‘Compound-Potential target’,‘Target-Disease’ and ‘Target-TCM syndrome’ networks.Finally,the compatibility mechanism of ‘Rhizoma Chuanxiong-Rhizoma Cyperi’ drug pair and the characteristics of "multiple components and multiple targets" in the treatment of qi stagnation and blood stasis syndrome were expounded.Its the potential pharmacological basis of Chaihu Shugan Powder to‘promote blood circulation and relieve pain’.(4)In Chapter 5,GC×GC-TOF/MS was used to separate and detect the volatile components of Chaihu Shugan powder,combined with ICMP-MSCC algorithm of stoichiometry method,and the difference of the volatile components of Chaihu Shugan powder and Sini powder was analyzed.The results showed that,the results of datas are consistent with the actual,can intuitively show the similarity and difference between different fingerprints.There are more medicinal substances and fragrance constituents of Chaihu Shugan powder.And the main volatile components are D-Limonene,Terpentene,Ligustilide,n-Butylphthalide,Butylphthalide,Vanillone,Naringone,Geronidene and Vanillone.Finally,the volatile components of Chaihu Shugan powder were predicted and screened,and 96 potential targets were associated with 11 volatile components.Among them,the volatile components n-Butylphthalide,Naringone and Butenylphthalide have higher connectivity and may plays important pharmacological activities.