A Preliminary Study On The Pharmacodynamic Material Basis And Mechanism Of Sichen Zhike Granules In The Treatment Of Acute Exacerbation Of Chronic Bronchitis

Sichen Zhike granules(SCZKG)is composed of Artemisia scoparia Waldst.et Kit.,Glycyrrhiza uralensis Fisch.,Phlomis brevidentata H.W.Li.and Bergenia purpurascens(Hook.f.et Thoms.)Engl.,which has the effect of "clearing heat and nourishing lung,relieving cough,relieving fever and nourishing vital energy".It is made by the classical prescription of Tibetan medicine theory.It has been proved by many years of clinical use that it is effective in the treatment of acute exacerbation of chronic bronchitis(AECB),however,its material basis and mechanism of action are not completely clear,and the overall quality of the compound needs to be evaluated.Combining the theories and methods of serum pharmacochemistry,molecular docking technology and network pharmacology,this paper intends to explore the pharmacodynamic material basis and mechanism of SCZKG in treating AECB and through the control of the content of each drug flavor index components,in order to evaluate the overall quality of the compound,and provide a basis for the further development and utilization of SCZKG.The content of this study mainly includes the following parts:1 Analysis of chemical constituents of SCZKG and its representative single herb based on UPLC-QE-Orbitrap-MS and UPLC-Q-TOF/MS techniquesUPLC-QE-Orbitrap-MS and UPLC-Q-TOF/MS have the advantages of fast analysis speed,high sensitivity and high throughput,so they are widely used in the component analysis of traditional Chinese medicine.In this experiment,based on UPLC-Q-TOF/MS combined with UNIFI screening platform for rapid identification and analysis of the chemical constituents of Bergenia purpurascens,a total of 43 chemical constituents were identified,of which 16 were found in Bergenia purpurascens for the first time.Based on the analysis of the chemical composition profile of SCZKG based on UPLC-QE-Orbitrap-MS,a total of 88 components were identified,including 54 from licorice,15 from Artemisia annua,9 from crab beetle and 12 from pakchoi.The study of chemical group in vitro lays a foundation for screening the in vivo components of SCZKG.2 Analysis of absorbed components in rat plasma of SCZKG based on UPLC-QE-Orbitrap-MS and UPLC-MS/MSIn this experiment,88 kinds of chemistry in SCZKG compound were identified based on UPLC-QE-Orbitrap-MS technology,but for biological samples,the matrix effect was great,and the ion inhibition phenomenon was obvious,which made it difficult for trace components in biological samples to appear in the spectrogram;UPLC-MS/MS adopted multi-reflection monitoring mode,which greatly improved the detection sensitivity and detection limit,and avoided the interference of complex background.Based on the high-throughput and rapid analysis of the chemical composition of SCZKG extract based on UPLC-QE-Orbitrap-MS technology,and combined with the targeted screening strategy of UPLC-MS/MS,the absorbed components in rat plasma of SCZKG were identified quickly and accurately.After oral administration,16 prototype components were identified in rat plasma,including 9 from licorice,4 from crab beetle,2 from pakchoi and 1 from Artemisia annua.These 16 ingredients will be used as potential pharmacological substances of SCZKG in the treatment of AECB.3 Screening of active substances of SCZKG in treating AECB based on network target analysis and molecular docking technologyIn this study,based on the absorbed components in rat plasma,the correlation between the absorbed components of SCZKG and AECB disease was analyzed,and the "core target-component" network was constructed to predict the material basis of SCZKG in treating AECB.The predicted active components were docked with the target of treatment of AECB,and the components with high matching degree were selected as potential pharmacological substances.The results showed that 94 common targets were obtained by mapping the targets of absorbed components in rat plasma with AECB disease targets,and the core targets of 20 components such as GAPDH,ALB and AKT1 were screened by PPI topological analysis.Through the construction of "core target-component" network,the potential active substances were preliminarily screened,and the top components were isoliquiritin,verbascoside,chlorogenic acid,liquiritin apioside,glycyrrhizin and so on.Sixteen kinds of absorbed components and core targets in plasma were screened by molecular simulation based on molecular docking technique.By comparing with the vina score value of dexamethasone,a positive drug for AECB,it was found that the binding of 8 components such asliquiritin and glycyrrhetinic acid was more stable than that of dexamethasone.We believe that the 8 ingredients that combine stably are the potential active substances.4 Prediction of the mechanism of SCZKG in treating AECB based on network pharmacologyIn order to explore the mechanism of SCZKG in treating AECB,the biological enrichment analysis and pathway prediction of 16 blood components of SCZKG were carried out by using network pharmacology technology.The results showed that 94 common targets were mainly involved in cell signal transduction(cell proliferation,cell migration,apoptosis),inflammatory reaction,angiogenesis and other biological processes,regulating PI3K-Akt,Rap1,Ras,TNF and other related signal pathways.Through the construction of the visual network of "compound-transitional components in blood-target-pathway",it is directly shown that SCZKG in treating AECB is the result of the synergistic effect of multi-components,multi-targets and multi-pathways.5 Determination of four components in SCZKG based on HPLCSCZKG is a new drug for the treatment of AECB.In order to control the overall quality of the compound preparation,a HPLC method for the determination of bergenin,shanzhiside methyl ester,chlorogenic acid and glycyrrhizin in SCZKG was established for the first time,which provides a scientific basis for the quality standard research of SCZKG.The results showed that bergenin,shanzhiside methyl ester,chlorogenic acid and glycyrrhizin showed a good linear relationship with the peak area when the contents of bergenin,shanzhiside methyl ester,chlorogenic acid and glycyrrhizin were 0.48?2.4 ?g,0.08?0.4 ?g,0.18?0.9 ?g and 0.12?0.6 ?g,respectively.The average recovery rate was in the range of 92-105%,indicating that the method is accurate and reproducible and can be used for the determination of SCZKG.