To Explore The Mechanism Of Dingchuan Decoction And Its Baicalin In The Treatment Of Heat Asthma Syndrome Inhibiting JAK/STAT6 Pathway

Part ?:Theoretical ResearchBronchial asthma defined in Western medicine belongs to the category of "wheeze" in traditional Chinese medicine.Due to abnormal congenital endowment and phlegm turbidity,Chinese medicine practitioners divide the disease stages of asthma into:onset period,chronic duration,Remission period.Asthma often occurs secondary to external sensations,improper diet,emotional disturbances,fatigue,sleepiness,etc.,phlegm obstructs the airway,and the lungs are subsided,and the patient will have sudden attacks of wheezing and wheezing.In the clinical onset of asthma,heat and wind syndrome are the most common.The Dingchuan Decoction discussed in this article is from the "She Sheng Zhong Miao Fang",which is a classic prescription for traditional treatment of hot asthma.Through multiple database searches,the author found that the clinical application of Dingchuan Tang in the treatment of hot asthma by experts from various regions is conclusive;and the main component of Dingchuan Tang,Scutellaria baicalensis,has also been shown to have anti-inflammatory and antiasthmatic effects.However,the mechanism and action target of Dingchuan Decoction are not clear,so based on the network pharmacology research,the possible action target of Dingchuan Decoction for treating asthma is first selected,followed by the selection of baicalin,the main monomer component of Scutellaria baicalensis,Studying its anti-inflammatory and antiasthmatic mechanism provides a basis for further revealing the action pathways and targets of Dingchuan Tang,and also lays a good foundation for the further development of new effective antiasthmatic compound.Part ?:Study on the effect target of Dingchuan Decoction on asthma based on network pharmacologyObjectiveTo establish the biological function and pathway network of Dingchuan Decoction's active components-target,protein interaction and target,and to explore the mechanism and target of Dingchuan decoction.MethodsThe active components of Dingchuan Decoction in the treatment of bronchial asthma were screened from the pharmacology database and information database of traditional Chinese medicine system.The screen criteria of ADME were:oral bioavailability? 30%,blood-brain barrier permeability?0.3,class drug? 0.18.The target protein information of the predicted active components of Dingchuan decoction was collected through the pharmacology database of traditional Chinese medicine system and transformed into target gene information by UniProt software.Target information of asthma was obtained by TTD,OMIM,GAD,drugbank,pharmacogenetics and PharmGKB database.The target of the active components of Dingchuan Decoction on asthma was obtained by the Cytoscape software.The drug target disease network,protein-protein interaction(PPI)network were constructed.The target of the active components of Dingchuan Decoction on asthma was analyzed by using David database,and the enrichment of the pathway of Dingchuan Decoction on asthma was analyzed by using KEGG.Results(1)76 active components of Dingchuan Decoction were collected and screened,and 100 target protein information of 76 active components of Dingchuan Decoction were collected through database.(2)52 potential targets of Dingchuan Decoction acting on asthma were obtained by analyzing the intersection of citoscape software and 542 asthma related targets.(3)According to GO analysis,Dingchuan decoction was involved in the biological process of bronchial asthma,including signal transduction,inflammatory response and immune function,protein degradation,protein phosphorylation.(4)Dingchuan decoction is involved in the main pathways of asthma,including PI3K Akt signaling pathway,ERK signaling pathway,Notch signaling pathway,TGF-b signaling pathway,mTOR signaling pathway,MEK-STAT signaling pathway,GAS-STINGT signaling pathway,cell receptor signaling pathway,NF kappa B signaling pathway,MAPK signaling pathway and Sonic Hedgehog signaling pathway.ConclusionDingchuan decoction is highly related to bronchial asthma.It can control the occurrence and development of bronchial asthma through multi-channel,multi-target and multi active substances.It has the potential functions of promoting signal transmission,inhibiting inflammatory reaction and promoting protein decomposition.Part ?:Inhibition and remodeling of Baicalin on allergic airway injury in miceObjectiveAsthma,as a chronic disease characterized by chronic airway inflammation and high responsiveness,seriously affects the quality of life of patients and increases the economic burden of society.Baicalin is a kind of traditional Chinese medicine with anti-inflammatory and anti-tumor effects,but its role and mechanism in asthma are not completely clear.This part mainly explores the inhibition and remodeling of Baicalin on allergic airway injury and inflammation in mice.MethodsFirstly,the asthma model of C57bL/6 mice induced by ovalbumin(OVAO)was established.The primary generation of inflammatory factors,protein expression and inflammatory cell population were detected by ELISA,Western blot and flow cytometry,respectively.Then,the asthmatic airway remodeling model of mice was made,and the rats were randomly divided into 6 groups according to body weight,namely blank group(control,n=10),model group(model,n=10),dexamethasone group(DS,n=10),baicalin low dose group(LD,n=10),baicalin middle dose group(MD,n=10),baicalin high dose group(HD,n=10)The expression of a-smooth muscle actin(a SMA),phosphorylated extracellular signal regulated kinase 1/2(pERKl/2)and transforming growth factor-? 1(TGF ? 1)protein in the lung tissue homogenate and the mRNA level of p-ERK1/2 in the airway smooth muscle(ASM)were detected by RT-PCR.In this study,SPSS 22.0 statistical software was used for statistical analysis.Results(1)Compared with the control group,the total cells,eosinophils,macrophages,lymphocytes and neutrophils of asthma mice increased significantly after ova inhalation.In addition,when baicalin was administered in a dose-dependent manner,the increase of the total number of the above cells and the number of the differentiated cells decreased significantly.(2)Compared with the control group,the RI value of ova stimulated group increased significantly after acetylcholine stimulation at different concentrations(2.5mg/kg,5mg/kg and 10mg/kg).In addition,baicalin decreased the increase of RI induced by ova in a dose-dependent manner,especially in the concentration of 40 mg/kg baicalin.Compared with the control group,the cdyn value of ova stimulation group decreased significantly.Baicalin or dexamethasone can effectively alleviate the decrease of cdyn induced by ova.(3)After sensitization,the levels of IL-4,IL-5,IL-6,IL-13 and TNF-? were significantly increased,but the levels of IFN-y had no significant difference among different treatment groups.Baicalin and dexamethasone can inhibit the expression of cytokines induced by ova.(4)Compared with the control group,the serum IgE and IgG1 levels of ova induced group were significantly higher.At the same time,baicalin or dexamethasone can effectively reduce the increase of serum IgE and IgGl levels after ova sensitization.However,there was no significant difference in serum IgG2a levels between the different treatment groups.(5)The levels of JAK,STAT6 and p-stat6 in the ova inhalation group were higher than those in the control group.More importantly,with the different concentration of baicalin or dexamethasone,the increased levels of JAK,STAT6 and p-stat6 sensitized by eggs will decrease in varying degrees.(6)In the blank group,airway epithelium was smooth,inflammatory cell infiltration was not visible,and tracheal wall was normal,In the model group,the bronchus became narrow and deformed,the bronchus closed and thickened,the mucosal plica increased,the inflammatory cell infiltration was obvious,and the phenomenon of accumulation appeared.Compared with the model group,the lung histopathology of the patients in dexamethasone group,Scutellaria baicalensis group and high and low dose group decreased,but the effect of dexamethasone group was the best.(7)Compared with the blank group,the expression of p-ERKl/2 protein in the lung tissue of the model group was significantly increased.Compared with the model group,the expression of p-ERK1/2 protein in the lung tissue of mice in the dexamethasone group and the high and low dose of baicalin decreased significantly.(8)The expression of ERK1/2 mRNA in the model group increased significantly compared with the blank group.Compared with the model group,the ERK1/2 mRNA expression in the dexamethasone group and the baicalin high and low dose groups was significantly reduced.Compared with the blank group,the expression of ?-SMA protein in the lung tissue of the model group was significantly increased.Compared with the model group,the expression of a-SMA protein in the dexamethasone group and the baicalin high and low dose groups decreased significantly.(10)The expression of TGF-?1 protein in the model group was significantly increased compared with the blank group.The expression of TGF-?1 protein in the dexamethasone group and the high and low dose groups of baicalin were significantly lower than that in the model group..Conclusions(1)Baicalin can relax the smooth muscle of mouse bronchus,resist the spasm and contraction of bronchus caused by acetylcholine and histamine,alleviate the symptoms of asthma and reduce the expression of inflammatory factors.It further revealed the molecular mechanism of baicalin inhibiting JAK/STAT6 pathway to reduce airway damage,which provided potential guidance for clinical treatment of asthma.(2)Baicalin may affect the synthesis of ?-SMA,ERK signaling pathway and ERK1/2 mRNA by reducing the expression of TGF-? 1 protein,so as to reduce airway inflammation,inhibit airway cytoplasmic fibrosis and improve airway remodeling.However,the specific molecular mechanism still needs further study.