Analysis And Research On The Role Of Yinhuang Qingfei Capsule Based On Integrated Pharmacology Strategy

Objective:Yin-Huang-Qing-Fei capsule(YHQFC),a classical traditional Chinese medicine(TCM)prescription consisting of 14 herbs,has shown broad clinical applications in chronic obstructive pulmonary disease and contains a variety of pharmacological compounds.However,the systematic analysis of chemical constituents of YHQFC is difficult to determine and thus remain unclear to date.At the same time,even though the YHQFC has been widely used in the treatment of chronic bronchitis,with good curative effects,the pharmacological mechanism of YHQFC remains unclear.To obtain the profile of the chemical compouns of YHQFC,and to explore the pharmacological mechanism of YHQFC acting on chronic bronchitis,the strategy of integrative pharmacology is applied to primarily analyze the effects of YHQFC.And this research will provide new view of study mode for TCM.Methods:A sensitive and efficient method based on ultra-high-pressure liquid chromatography coupled with linear ion trap-Orbitrap tandem mass spectrometry(UHPLC-LTQ-Orbitrap)was developed to characterize the chemical constituents of YHQFC;the putative target profile of YHQFC was predicted using MedChem Studio,based on structural and functional similarities of all available YHQFC components to the known drugs obtained from the DrugBank database,the common targets between any two of herbs contained in YHQFC was calculated;to observe the relationship between the efficacy and the modern pharmacological activity of a herb,enrichment analysis of putative targets of each herb was performed based on the theory of Chinese medicine,and a herb-efficacy-pharmacological activity-parthway network was constrcted;known therapeutic targets of chronic bronchitis was collected to build putative targets of YHQFC-known therapeutic targets of chronic bronchitis network.Following the calculation of four topological features(degree,node-betweenness,closeness,and coreness)of each node in the network,major putative targets were identified and used to perform pathway enrichment analysis.Then the herb-target-pathway network was constructed to define the biological processes related to YHQFC actiong on chronic bronchitis.The key chemical compounds were identified according to the crucial targets of YHQFC.Finally,a molecular docking simulation was performed to observe the binding capacity between the chemical compounds and targets to validate the reliability of the prediction of network pharmacology.Results:1.Based on mass spectrometry data,chromatographic behaviors,reference standards and previous literatures,a total of 204 compounds were unambiguously or tentatively characterized,mainly including 68 flavonoids,25 alkaloids,22 lignans,22 organic acids,17 triterpenoids,11 steroid saponins,11 coumarins,6 terpene trilactones,5 limonins and 17 other compounds.The sources of these compounds were also illustrated.37 ingredients from Glycyrrhiza uralensis Fisch.(GC)were identified,while 29 ingredients were from Ginkgo biloba L.(YXY),26 ingredients were from Citrus aurantium L.(ZS),24 ingredients were from Schisandra chinensis(Turcz.)Baill.(WWZ),15 ingredients were from Eriobotrya japonica(Thunb.)Lindl.(PPY),14 ingredients were from Fritillaria thunbergii Miq.(ZBM),13 ingredients were from Artemisia rupestris L.(XJYZH),12 ingredients were from Isatis indigotica Fort.(DQY),12 ingredients were from Dioscorea nipponica Makino(CSL),10 ingredients were from Ephedra sinica Stapf(MMH),10 ingredients were from Semen Armeniacae Amarum(KXY),8 ingredients were from Acorus tatarinowii Schott(SCP),6 ingredients were from Lepidium apetalum Willd(BTLZ).2.A total of 26917 pairs of chemical compounds and targets were predicted,and there were different numbers of common putative targets among the herbs contained in YHQFC,suggesting that these herbs might have several interactions in the course of treatment.Moreover,.50%of the putative targets of PPY were common to those of the other herbs,implying a close relationship between PPY and other herbs in the formula.3.According to the herb-efficacy-modern pharmacological activity-pathway network,the efficacy has a close relationship with modern pharmacological activity of the same herb.As for the putative targets of YHQFC and known therapeutic targets of chronic bronchitis network,a total of 475 crucial targets were selected and applied to prform enrichment analysis.It is suggested that that the major putative targets of YHQFC are mostly involved in pathways related to anti-inflammation processes and the immune system,which may be crucial elements during the attenuation of chronic bronchitis.Besides,we also found that YHQFC may exert its therapeutic effects on chronic bronchitis by regulating the pathways related to the contraction of airway vascular smooth muscle.The relevant nerve center regulation induced by the major putative targets of YHQFC was another pathway to calm cough and asthma.Eight major targets including IL-3,IL-5,the FcεRI receptors[FCER1G],eosinophil peroxidase[EPX],IL-4,IL-10,IL-13,and a member of the eotaxin subfamily of chemokines[CCL11]were demonstrated to participate in the asthma pathway,which has been shown to be involved in the occurrence and progression of chronic bronchitis.As shown in Figure 3,the asthma pathway is mainly composed of inflammatory processes.According to the clusters of major hubs contained in the asthma pathway,YHQFC may be of vital importance for regulating these cytokines and chemokines.And the molecular docking simulation results indicated that 17 pairs of chemical components and candidate YHQFC targets involved in asthma pathway had strong binding efficiencies.Conclusion:A LC-MS/MS method was developed to characterize the chemical compounds of YHQFC,and 204 constituents were identified.This would provide evidence for quality control and pharmacological studies of YHQFC.Network pharmacology technique was utilized to predict putative targets and construct relevant networks to elaborate the pharmacological mechanism of YHQFC acting on chronic bronchitis at a computional level.Features and innovation points of this paper are as follows:1.A total of 204 chemical compounds of the big formula Yin-Huang-Qing-Fei capsule were identified through LC-MS/MS based on the strategy of integrative pharmacology;2.The network of interactions among crucial chemical compounds of the big formula Yin-Huang-Qing-Fei capsule,targets and pathways was constructed to elucidate the molecular mechanism of Yin-Huang-Qing-Fei capsule acting on chronic bronchitis based on network pharmacology.The molecular docking analysis was performed to validate the binding efficiencies of active chemical compounds of YHQFC combined with main targets involved in asthma pathway.