Study Of The Establishment Of ACO Disease Animal Model And The Discovery Of Its Potential Mechanism Targets And Candidate Drugs

Asthma-chronic obstructive pulmonary disease overlap(ACO)is a severe clinical syndrome,which refers to patients with both asthma and COPD disease clinical features.Compared with simple asthma or COPD,ACO patients have more severe clinical symptoms,more frequent attacks,a more significant decline in quality of life,and a significantly shorter survival time,which poses a serious threat to the quality of life and life safety of patients.Moreover,the medical occupancy rate and medical expenses of ACO patients have risen sharply,which also brings a heavy burden to the social economy and medical resources.However,ACO has many difficulties and uncertainties in its clinical diagnosis and treatment,and the screening and evaluation of its therapeutic drugs have not yet started,especially since its disease animal model has not been fully established,which has seriously affected the discovery of its disease target mechanism and effective drugs.Therefore,based on the clinical disease characteristics of ACO,the establishment of animal models of ACO disease and the exploration of the common and different pathological mechanisms of ACO,asthma,and COPD are particularly important for the development of targeted therapeutic drugs.In response to these,this thesis firstly took ACO clinical disease characteristics as the guidance,use OVA,LPS,cigarette smoke,and other asthma and COPD model conditions to induce the establishment of the ACO inflammation mouse model.Further used RNA-seq detection technology to explore the difference in gene expression in lung tissues of ACO,asthma,and COPD mice have revealed the potential mechanism targets of ACO inflammation.Simultaneously,based on the extensive research of natural source compound resveratrol in treating diseases,our team's early discovery of its dimer-derived compounds had good efficacy in asthma and CPD airway inflammation research.Using network pharmacology methods,mining literature data of resveratrol disease targets,constructing resveratrol-disease-target network,discovering potential targets of this kind of drugs,and combining them with the results of RNA-seq omics research explored a potential target for this class of compounds in the mechanism of ACO.Based on this,the team had newly synthesized 3-arylbenzofuran derivative EAPP-2 by referring to the chemical structure characteristics of resveratrol dimer,the preliminary study of its effect on the treatment of ACO inflammation and its potential mechanism targets.The main research results are summarized as follows:1.Based on the clinical characteristics of ACO,using OVA,LPS,cigarette smoke,and other asthma and COPD model conditions superimposed,established a mouse model consistent with the necessary clinical characteristics of ACO.Model mice showed overlapping features of asthma and COPD in lung histopathological changes and levels of inflammatory factors in bronchoalveolar lavage fluid,which better mimic some of the clinicopathological features of ACO.This model can provide a tool for studying the pathological mechanism of ACO and the screening of potential drug candidates.2.Using RNA-seq detection technology,the differences in gene expression in lung tissues of ACO mice were found.The results of the study found potential 6324 differentially expressed genes in ACO mice,including 2717 down-regulated genes(42.7%)and 3607 up-regulated genes(57.3%),with pneumonia,pulmonary fibrosis,chronic obstructive airway disease,lung tumors,rheumatoid arthritis,and other gene profile characteristics are in line with the clinical features of ACO.Moreover,the underlying molecular mechanisms of ACO lung inflammation and fibrosis were mainly related to the HLA-DRA,SYK,CTLA4,VAV1,NRAS,and JAK3 signaling pathways.This study provides a basis for studying the mechanism of ACO and its drug discovery.3.Based on the data reported in the literature on potential targets of resveratrol disease treatment,using network pharmacology methods,the resveratrol-disease-target network was constructed,and the critical signal pathways and molecular mechanisms were explored through GO enrichment analysis and KEGG pathway analysis.Signal pathways and molecular mechanisms have been discovered related to targets such as STAT3,AKT1,SRC,JAK,and VEGFA,and multiple signaling pathways such as T cell receptors,MAPK,Toll-like receptors,Fc?R1,and TNF.This study uncovered potential targets of stilbene compounds such as resveratrol in respiratory diseases and provided enlightenment to modify and optimize these compounds and their development research in respiratory diseases.4.Based on the above research results,the anti-ACO airway inflammation effect and mechanism of 3-arylbenzofuran derivative EAPP-2 were studied.The results showed that EAPP-2 could significantly inhibit the inflammatory characteristics of ACO mice,reduce the infiltration of pneumonia cells,improve the inflammatory pathological changes in lung tissue of ACO mice,and reduce the fluidity of inflammatory cells inflammatory factors in bronchoalveolar lavage fluid.It could reduce the effect of general IgE and IgG1 products in peripheral blood and effectively inhibit the inflammatory response of RAW264.7 cells induced by LPS.Moreover,EAPP-2 could significantly reduce the expression and phosphorylation of Syk,NF-?B,as well as expression of NLRP3 and iNOS in the lung tissue of mice with ACO airway inflammation and LPS-induced RAW264.7 cells.The results suggested that the anti-ACO inflammation effect of EAPP-2 might partly be achieved by inhibiting the expression and phosphorylation of Syk,thereby inhibiting the NF-?B and NLRP3 pathways.This study provides a preliminary basis for the development of EAPP-2 in ACO treatment and can reference the discovery of ACO treatment drugs targeting Syk.