| Lung cancer is now one of the cancers with highest incidence and damages the health of human seriously.In the previous study of our research group,the Jinfukang prescription was optimized by prescription optimization and results of previous studies showed that Astragalus plays an important role in the Jinfukang optimization prescription's anti-lung cancer activity.In order to clarify the material basis of Astragalus,and the quantity and quantity ratio relationship of composition,we employ various technologies and methods to explore the ultimate problem.This paper focuses on five parts and the specific content is as follows:The components prediction of Astragalus' material basis of anti-lung cancer effect through multi-target and multi-pathway:The network of components-function channel-target was built through information retrieval and text analysis.Then,the potential active components,function pathways and targets were predicted according to the information of network topology.After that,the 100mg/kg benzopyrene was injected intraperitoneally to build the situ lung cancer model.The lung tissues in blank,model and high dose group were collected for the following microarray experiment.The result revealed that benzopyrene could induce the disorder of gene networks in lung tissues of normal AJ mice in the way of up-regulating and down-regulating target genes.However;Astragalus membranaceus could make the aberrant expression of genes return to normal range and correct the pathological disorders caused by benzopyrene.Based on the results of network pharmacology and gene chips experiments,we predicted that Astragalus saponins including Astragaloside ?,Astragaloside ?,cycloastragenol,Astragaloside VII,Astragaloside I and Astragalus flavonoids including calycosin,formononetin,biochanin A,genistin,Calycosin-7-O-?-glucoside,Ononin,Daidzein are components which played important roles in the prevention activity on lung cancer.The finding of Astragalus' material basis components of prevention effect on lung cancer in the autophagy pathwayCigarette smoke was used to induce the occurrence of vitro lung cancer of NHBE cells.After the administration,Western Blot,Immunofluorescence and PCR were utilized to investigate the expression level of relative signal proteins and mRNA and clarify the underlying mechanism.Besides,UPLC/TOF/MS was also employed to examine the components of ethanol extract of Astragalus.The vitro experiments have shown that Astragalus membranaceus has favorable preventive effect on lung cancer and it may play the role through the regulation of cell autophagy.In addition,8 components have been qualified including Calycosin-7-O-?-glucoside,Ononin,calycosin,formononetin,Astragaloside ?,Astragaloside IV,cycloastragenol and Astragaloside I.As a result,the 8 components were identified as the components of material basis of Astragalus' prevention effect on lung cancer in the autophagy pathway.The impact of metabolism on the quantity and quantity ratio of Astragalus'material basis components of preventive effects on lung cancerLiquid chromatography tandem mass spectrometry was used to determine the metabolites change qualitatively and quantitatively and to reveal the potential rules.The results showed that many constitutes in Astragalus membranaceus have experienced metabolism and transformation and the main metabolic pathway is hydrolysis,which converts the prototypes into metabolic products with lower polarity and smaller molecular weight.Before incubation,the ratio of components was Calycosin-7-O-?-glucoside:Ononin:calycosin:formononetin:Astragaloside ?,Astragaloside ?:cycloastragenol:Astragaloside I=1550.1:587.2:250.2:138.3:2460.0:1180.3:200.2:27.6.After 72h incubation,however,the ratio of components was Calycosin-7-O-?-glucoside:Ononin:calycosin:formononetin:Astragaloside ?,Astragaloside ?:cycloastragenol =10.3:7.3:45.1:267.9:12.2:67.8:706.1.Besides,there was difference between the metabolic conversion ratio of different components,which shows the programmatic releasing characteristics of the Chinese medicine preparation.Thus,it is important to design reasonable release unit to realize the long-time,reasonable and programmatic release of the preparation.Screen the bioactive preventive lung cancer components with cell solid chromatography modelA549 cell line was utilized for the building of the cell solid chromatography model.The A549 cell was incubated with metabolites obtained from the previous experiment.The intracellular fluid and dissociation solution were collected for the HPLC,UPLC/MS/MS and UPLC/TOF/MS analysis after the sample pretreatment.Among these analysis techniques,HPLC was used for the optimization of incubaition condition including incubation concentration,incubation time and elution times of PBS.The results revealed that the optimal washing times is 8 and the incubation concentration and time are 2×10-4g/ml and 120min,correspondingly.The results of UPLC/MS/MS and UPLC/TOF/MS analysis showed that 6 compounds including Calycosin-7-O-?-D-glucoside?Ononin?Calycosin?Astragaloside ??Cycloastragenol as well as the metabolic product of Astragaloside ?,the code of which was C7 in this paper,could combine with the cell membrane.In addition,3 compounds including the reduction product of Calycosin,the code of which is C13 in this paper,Calycosin and Formononetin could combine with the intracellular protein after passive transport across the cell membrane.In conclusion,all of the 8 components comprised the material basis of preventive effect of Astragalus membranaceus on lung cancer.Study on biopharmaceutical properties of Astragalus saponins and Astragalus flavonoidsAccording to the results of intestinal flora and cell solid chromatography experiments,we will focus on the investigation of the biopharmaceutical properties of flavonoids and saponin components and aim to clarify the solubility and permeability of them for the guidance of choosing proper preparation form and maximum of drug effects.The results showed that according to the equilibrium solubility,saponin should be divided into 2 sub-components.Specifically,Astragaloside IV and Astragaloside ? were divided into sub-component a and Cycloastragenol was divided into sub-component b;The flavonoids were also should be divided into 2 sub-components and Calycosin-7-O-?-D-glucoside as well as Ononin was deemed as sub-component a while Calycosin and Formononetin was deemed as sub-component b.According to the results of apparent oil-water partition coefficient in water and different pH value of solution,all the components in saponin and flavonoids shared similar properties in the aspects of solubility and oil-water partition coefficient.Then,the global property was obtained using weight coefficient method.The results indicated that both the saponin and flavonoids components were characterized with poor solubility and good absorptivity and they should been classified to II type traditional Chinese medicine components according to BCS classification systems. |
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