Study On The Pharmacodynamics Of Characteristic Components Of Honey-processed Astragalus Based On Metabolomics

Background:The theory of traditional Chinese medicine believes that honey-processed astragalus can enhance its tonify Qi effects and Yang effects,thus is utilized to replace astragali radix in the spleen and lung Qi deficiency,the depression of mid-Qi syndrome.In our previous pharmacodynamics study,the results showed that the honey-processed astragalus could improve the physical signs,physiological and biochemical indexes of the spleen Qi deficiency better than the raw astragalus.The astragaloside IV,astragalus I and isoflavones glucoside was determined as the characteristic components of honey-processed astragalus on basis of the chemical characterization of compounds in raw and honey-processed Astragalus.In this study,we applied the metabolomics combination with network pharmcology for evaluating the pharmacodynamics of characteristic components in honey-processed astragalus and exploring its mechanism.Methods:Spleen Qi deficiency rat model was established by fatigue and restricted diet.Application the physiological and biochemical indexes in rats for investigating the efficacy of astragaloside IV,astragalus I and isoflavones glucoside.The serum and urine metabolomics base on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)and nuclear magnetic(~1H-NMR)were applied to monitor the biomarkers and pathways related to effect of pharmacodynamicsandtoevaluatethepharmacodynamicsof honey-prosecced astragalus.Network pharmacology was used to screen potential targets of characteristic components of honey-prosecced astragalus and to discusse its potential mechanism.Results:(1)Lymphocyte count,leukocyte count,erythrocyte number,hemoglobin content,serum d-xylose absorption value and physical sign score significantly decreased in spleen qi deficiency rats(P<0.05),which showed that the spleen qi deficiency rat model has been constructed successly.(2)After spleen Qi deficiency rat treatment with astragaloside IV,astragalus I and isoflavones glucoside,the above indexes significantly were improved and the number of lymphocyte,white blood cell,erythrocyte and hemoglobin content were increased.Meanwhile,the expression of cytokines IL-6 and TNF-αwas decreased and IFN-γwas increased in rats with spleen qi deficiency.Astragaloside IV inhibits the expression of TNF-αand IL-6 better than other characteristic components,and can promote the expression of IFN-γ.(3)18 potential biomarkers related to pharmacodynamics were screen out from serum its mainly phospholipids and choline substances such as lyso-phosphatidylcholine and phosphatidylcholine,and 23 potential biomarkers were screen out from urine its mainly amino acids substances contains serine,creatinine,hippuric acid,uric acid,etc.These biomarkers primarily involved in tryptophan metabolism,the tricarboxylic acid cycle,phenylalanine metabolism,arachidonicacidmetabolism,glycerolphospholipid metabolism and amino acid metabolism.Interestingly,the characteristic components of honey-prosecced astragalus can recover the content of the above biomarkers and the trend was close to the normal group in which honey-prosecced astragalus was closest to the normal group,followed by isoflavone glycoside and astragaloside IV.(4)The potential targets of 29were screened by the network pharmacological analysis,including 7 of astragaloside IV,10 of the isoflavones glucoside,and 12 of astragaloside I.These targets are including MIF,ALDH2,AMH,DNMT1,MAPK14 and so on.The target pathways analysis showed that these targets were related to protein synthesis,energy metabolism,tryptophan metabolism,inflammatory reaction and lipid metabolism.Conclusion:The characteristic components of honey-prosecced astragalus can improve the spleen qi deficiency and improve the immune function of spleen qi deficiency rats by regulating the immune response of cytokines.The mechanism is that regulation of arachidonic acid metabolism and glycerol phospholipid metabolism to improve the immune function,regulation of tryptophan metabolism,tricarboxylic acid cycle and amino acid metabolism to promote energy supply and protein synthesis.Targets of isoflavone glycosides were mainly involved in protein synthesis,energy metabolism and purine metabolism.Targets of astragaloside IV and astragaloside I were mainly involved in inflammation and amino acid metabolism.UPLC-Q-TOF/MS and ~1H-NMR based metabolomics combined with network pharmacology and bioinformatics analysis method provided a new insight for the study of pharmacodynamics of honey-prosecced astragalus and its characteristic components.