Effective Constituents And Mechanism Study Of Shengxian Decoction In The Treatment Of Chronic Obstructive Pulmonary Disease Based On Network Pharmacology And Metabolomics

The history of Shengxian decoction(SXT)is listed in the Intergrating Chinese and Western Medicine,which consists of five herbs including Astragali Radix,Bupleuri Radix,Cimicifugae Rhizoma,Anemarrhena asphodeloides Bge and Platycodonis Radix,SXT is widely used in the treatment of heart and lung diseases.Modern pharmacological studies have demonstrated the good therapeutic effect of SXT on chronic obstructive pulmonary disease(COPD),but the key effective constituents and mechanism of action have not been fully elucidated.Due to the characteristics of multi-component Chinese medicine acting on multiple targets,the traditional"point-to-point"research method is difficult to screen the effective constituents and elaborate the mechanism of action of multi-component Chinese medicine acting on multiple targets from a comprehensive perspective.The holistic approach based on metabolomics and network pharmacology is consistent with the holistic concept promoted by Chinese medicine research,which not only allows the study of the mechanism of action of complex systems of Chinese medicine from multiple perspectives,but also allows the screening of key effective constituents.Therefore,in this study,based on the clarification of the treatment effect of SXT in animal models of COPD,the endogenous biomarkers affected by the disease and the modulating effect of SXT were evaluated using metabolomic methods of serum and urine samples.By constructing a metabolic regulatory pathway map for the treatment of COPD with SXT,the mechanism of action of SXT in the treatment of COPD was elucidated at the metabolic level.In addition,based on comprehensive characterization of the chemical constituents of SXT in vitro and the blood constituents in vivo,the pharmacology of the targeting network was used to explore the key pharmacological components,core targets and related metabolic pathways.The molecular docking technique was used to validate the screened key pharmacological components and core targets,to elucidate the pharmacological components and mechanism of action of SXT in the treatment of COPD at the molecular level.The main research contents are as follows1.pharmacodynamic evaluation of SXT in the treatment of COPDCigarette smoke combined with intratracheal lipopolysaccharide(LPS)drip method was used to establish a COPD rat model and evaluate the therapeutic effect of SXT in the treatment of COPD.The results showed that SXT can significantly regulated the body weight of COPD rats;reduced the damage of lung tissues and small intrapulmonary airways;decreased the alveolar diameter,mean lining interval and thickness of small intrapulmonary airways;increased the mean alveolar number;decreased the levels of inflammatory factors IL-1βand IL-6;increased SOD and decreased MDA level;and decreased the expression of TNF-αand TGF-βin the lung tissues of rats.The above results demonstrate that SXT acts as an anti-COPD effect by improving body weight,lung tissue damage,inflammatory response,and oxidative stress damage in rats with COPD.This section lays the foundation for the study of the mechanism of action of SXT in the treatment of COPD.2.Study the mechanism of action of SXT in COPD based on metabolomicsBased on the UPLC-Q-TOF-MS metabolomics method,we comprehensively analyzed the changes of urine and serum metabolic profiles in COPD rats,and explored the mechanism of action of SXT in COPD from the metabolic level.The results identified 44 biomarkers(25 in urine,20 in serum,and 1 significantly different in both serum and urine)closely associated with the occurrence and the development of COPD,of which 42 were significantly regressed by SXT.The results of metabolic pathway analysis showed that the biomarkers mainly acted on metabolic pathways such as arachidonic acid metabolism,primary bile acid biosynthesis,TCA cycle,taurine and hypotaurine metabolism,tryptophan metabolism and unsaturated fatty acid biosynthesis.Based on the analysis of the biological significance of the metabolic profile,the treatment effect of SXT on COPD may be achieved through the regulation of abnormal amino acid metabolism,imbalance of intestinal microbiota and energy metabolism,and inhibition of inflammatory damage.The endogenous component changes perturbed by COPD and the mechanism of action of SXT were described at the metabolic level.3.Analysis chemical constituents of SXTThe UPLC-IMS-QTOF-MS technique was used to detect the SXT extract in both positive and negative ion modes.DDA,DIA and HDMS~E three data acquisition modes and UNIFI software were used to assist the analysis.As a result,a total of 120compounds in SXT were identified or tentatively characterized,including 74 saponins,17 flavonoids,7 cinnamic acid derivatives,8 triterpenoids and 14 others.Furthermore,high-definition MS~E was used to identify six pairs of co-eluting isomers from SXT.This study laid the material foundation for the screening of the effective constituents of SXT and the study of the mechanism of action.4.Study on the effective constituents and mechanism of action of SXT based on targeted network pharmacologyBased on the chemical constituents analysis of SXT in vitro,UPLC-Q-TOF-MS technique was used to analyze the constituents in the blood.Combined with targeted network pharmacology and molecular docking techniques to further investigate the key effective constituents and core targets of SXT.With the help of UNIFI software,a total of 27 blood entry prototype constituents were identified,including saponins,flavonoids and cinnamic acid derivatives.A targeted network pharmacology approach was used to construct the"blood-entry constituents-disease target"network of SXT.The results suggest that the key effective constituents,such as ononin,formononetin,calycosin and moupinamide,may act on TNF,VEGFA,SRC,EGFR,JUN and STAT3 to moderate the PI3K-AKT and EGFR signaling pathways to treat COPD.The molecular docking results showed that each ligand molecule bound well to the target receptor.The above experimental results elucidate the key effective constituents and mechanism of action of SXT in the treatment of COPD.In conclusion,we have taken the lead in using network pharmacology and metabolomics technologies to comprehensively analyze the material basis and mechanism of action of SXT in the treatment of COPD at the molecular and metabolic levels.In addition,UPLC-IMS-QTOF-MS technology combined with DDA,DIA and HDMS~E data acquisition modes were used for rapid and systematic analysis of the chemical constituents of SXT for the first time.Our study provides a new method for the screening of effective constituents and mechanism of action of traditional Chinese medicine,and provides data support for further development and utilization of SXT.