| Chinese Medicine Formula is the main application form of Traditional Chinese Medicine in curing disease,and it's the directly manifests of Chinese medicine compatibility theory.Due to its characteristics like complex composition,multiple targets and the unclear medicinal ingredients,the modernization process of Chinese Medicine has been seriously restricted.Therefor,analyze the the chemical composition of Chinese Medicine,screening the pharmacodynamic substance and investigate the mechanism in vivo through modern instrumental analysis techniques is an important method which promote the modernization of traditional Chinese medicine.FuFang-Xialian-Capsule(FXL)astheimprovedformulaof Banxia-Xiexin-Decetion,there is no report about it.It means that,the chemical composition,medicinal ingredients and drug target are still unclear,the metabolic process of FXL in vivo is not known yet.In this paper,the composition of FXL was investigated for first time,and then the pharmacodynamic substance and drug targets could be analyzed according the results of composition analysis.Finally,the metabolite process of pharmacodynamic substance in vivo could be clarified.The research content mainly includes the following aspects:1.The chemical constituent's analysis of FXLAn ultra high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UHPLC-Q-TOF-MS)method and an ultra high-performance liquid chromatography coupled with Linear Ion Trap Mass Spectrometer(UHPLC-IT-MS)method were appliacate to analyze the constituens of FXL.UPLC-Q-TOF-MS could provide the accurate mass of each compound(within10 ppm)and UPLC-IT-MS could acquired the fragment ions information of each compound,combine the data acquired and compare with the previous reports and the mass data acquired from standard substance,a total 73 compositions were identificated.The componsitions include 8 alkaloids,30 flavonoids,21 triterpenes,11 ginsenosides,1 coumarine and 2 lignins.The source of alkaloids were Coptis Chinensis and Mangnolia officinalis,the source of triterpenes and coumarine was Radix Glycyrrhizine Preparata,the source of ginsenosides and lignins were Panax Ginseng and Mangnolia officinalis indeivdualy.The results above provide the basis for the further research about the screening of medicinal ingredients and the mechanism research in vivo.2.The antioxidant activity of FXLThe damage of oxygen free radical to gastric mucosa is one of the important pathogenesis which caused chronic atrophic gastritis.Therefore,evaluate the radical activity of scavenging free radical is very important to evaluate the drug activity and the mechanism of drug action.In this paper,FRAP assay and DPPH free radical scavenging assay were utilized to evaluate the antoxidant activity of FXL,the formula ingredients and the main activity compounds in FXL.The results showd that FXL has a moderate antioxidant activity,the formula ingredients such as Mangnolia officinalis,Scutellaria baicalensis and Coptis Chinensis have good antioxidant activity.It means that Mangnolia officinalis,Scutellaria baicalensis and Coptis Chinensis are the main formula ingredients which play antioxidant activity in FXL.In order to further elucidate the main antioxidant activity components in FXL,the compontes which from these three formula ingredients were selected to screen the antioxidant activity components.The results showed that acteoside,isoacteoside,baicalin and baicalein have good antioxidant activity,these four components may the main antioxidant activity components of FXL.The components from Coptis Chinensis don't showed excellent antioxidant activity;it means that the components from Coptis Chinensis may play antioxidant activity through synergy.This section elucidates the main pharmacodynamic substance and potentinal mechanism of FXL from the view of oxidative stress.3.The pharmacodynamic substance and pharmacologic mechanism of FXL in chronic atrophic gastritisThe pharmacodynamic substance of drugs must be the components which could absorbed in vivo and reach the targets organs.In order to screening the pharmacodynamic substance and pharmacologic mechanism of FXL in chronic atrophic gastritis,a new network pharmacology approach which was based on the components absorbed into blood was utilized.First,UPLC-Q-TOF-MS method was utilized to investigate the components absorbed into blood of FXL,a total 22components include 7 alkaloids,9 flavonides,5 ginsenosides and 1 triterpenes were absorbed into blood.Then network pharmacology approach was carried out according to the results of components absorbed into blood.Compound-compound target,pharmacodynamic substance-action target and action target-pathway network were built.Through network pharmacology approach,the relationships of pharmacodynamic substance-action target-pathway were analyzed.Finally;the pharmacodynamic substance and the pharmacologic mechanism of FXL in chronic atrophic gastritis were explored.4.The metabolite identification of FXL in vivoThe drug which absorbed in vivo could form a series of metabolites through metabolic reactions;the process of the metabolites formed is the metabolic process of drug in vivo.In order to investigate the metabolic mechanism of FXL in vivo,a Target-Group-Change strategy was founded and applied to the metabolite identification of FXL in vivo.UPLC-Q-TOF-MS~E method was utilized detected the metabolites in rat's plasma,and then analyze the data under the Target-Group-Change strategy.As the results,the prototypes which could transform into metabolites through metabolic reaction include magnoflorine,berberrubine,berberine,jatrotthizin,palmatine,baicalin,5,6,7-trihydroxy-8-methoxyflavone-7-o-glucuronide,wogonoside,wogonin,Ginsenoside R_f and Ginsenoside R_d,and there were 33 metabolites transformed from these prototypes were identificated.Based the results above,the network diagram of the metabolic relationship of them was inferred,the metabolic mechanism of main pharmacodynamic substance in FXL was preliminary elaborated.5.The pharmacokinetic study of FXLIn order to investigate the time-effect relationship of pharmacology substance after oral FXL in vivo,a rapid,accurate and sensitive UPLC-MS/MS method which could measure magnfolorine,berberine,jatrorrhizine,palmatine,glycyrrhizine,wogonoside,baicalin and wogonin at the same time were founded and applied to the pharmacokinetic study of FXL.As the results,the drug-time curve and pharmacokinetic pharameters were acquired.Combine the results of pharmacokinetic and results of metabolic in vivo,the reason for the change of serum concentration could be explained. |
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