Study On The Material Basis And Quality Evaluation Method Of Sheng-Yang-Yi-Wei Decoction

The classic formula Sheng-Yang-Yi-Wei decoction(SYYWT)is composed of 14 herbs including Astragalus propinquus,Glycyrrhizae preparata,Panax ginseng,Rhizoma pinelliae,Radix Paeoniae Alba,Saposhnikovia divaricata,Notopterygium incisum,Angelica biserrata,Pericarpium Citri Reticulata,Alismatis Rhizoma,Poria cocos,Radix Bupleuri,Macrocephalae Rhizoma,and Coptidis Rhizoma.which is used to treat symptoms of the spleen and stomach and their abnormal functions resulting in physical lethargy,etc.It is remarkably effective and widely used.However,Although SYYWT has been applied in many experimental studies of clinical diseases,the current material basis research and quality standards are not clear,and it fails to meet the requirements of the guidelines for the development of classical famous prescriptions.In this thesis,we analyzed the in vitro and in vivo chemical composition of SYYWT,and used targeted network pharmacological research methods to initially screen the pharmacodynamic substance bases acting in SYYWT and increase its quality control index.The fingerprints of SYYWT were performed by high performance liquid chromatography(HPLC)analysis and the content determination method of the pharmacological indexes,and then the quality evaluation criteria of SYYWT were improved.The main study of this research is as follows:1.Analysis of chemical constituents of SYYWT in vitroThe aqueous extract of SYYWT was examined by UPLC-Q-TOF-MS~E technique in positive and negative ion mode,and the in vitro chemical composition of SYYWT was analyzed with the aid of UNIFI software.A total of 120 compounds were identified in the aqueous extract of SYYWT by comparison with the standards combined with secondary tandem fragmentation information discrimination.They were mainly alkaloids,saponins,coumarins,flavonoids,terpenoids,organic acids and other components,which initially clarified the chemical composition of SYYWT aqueous extract.2.Analysis of blood components of SYYWT and pharmacological study of targeting networkFirstly,UPLC-Q-TOF-MS~E method and UNIFI system were applied to analyze the blood components of SYYWT,and then the"blood composition-target-disease"network of SYYWT was constructed with the help of targeting network pharmacological research strategy to investigate the main pharmacological constituents and key targets.It was found that a total of 27 prototype ingredients of SYYWT could enter the bodies of rats.It is speculated that these compounds may be the main pharmacological components of SYYWT.The"blood-entry component-target-disease"correlation network constructed by network pharmacology indicated that seven blood-entry components,including berberine hydrochloride,might act on STAT3,SRC,HSP90AA1,PIK3CA,MAPK1,MAPK3,EGFR,AKT1,VEGFA and other key targets in gastritis.The therapeutic effect was exerted.Therefore,it was tentatively determined that the seven blood components such as berberine hydrochloride may be the key components of SYYWT to exert its medicinal effects,which can be used as a reference basis for screening the quality standard index components of SYYWT.3.Study on HPLC fingerprint of SYYWTThe liquid phase spectrum of SYYWT was systematically optimized by high performance liquid chromatography(HPLC).15 batches of SYYWT were examined,and the common peaks were screened,and the fingerprint analysis method of SYYWT was successfully established after the attribution of common peaks and methodological investigation.After comparing the results of chemical constituents in vitro,20 common peaks were identified in the fingerprint of SYYWT and similarity analysis was performed.The data results indicate that the method is stable and feasible,and this fingerprint study can help to establish and improve the quality standard of SYYWT.4.Study on the determination of the concentration of SYYWTCombined in vitro-in vivo chemical composition analysis and targeted network pharmacology study to determine the quality standard index components of SYYWT Two content determination methods were established using different detectors according to the differences in chemical properties among different components:The content determination methods of five components,including berberine hydrochloride,were established by HPLC-UV method.The content determination methods of ginsenoside Rb1 and astragaloside were established by HPLC-ELSD method.After verification,the above two methods are stable and feasible,and can be used as the quality evaluation method of SYYWT.In conclusion,this paper initially screened the index components of SYYWT through in vivo and ex vivo chemical composition analysis combined with targeted network pharmacology,on this basis,the material basis and quality evaluation methods of SYYWT were completed,which provides a reference basis for the comprehensive quality control of the classical formula SYYWT and subsequent related research.