Studies On The Pathogenesis Of Asthmatic Phenotypes Based On Multi-omics And Therapeutic Strategy With The Combination Of Rhodiola Wallichiana Var.Cholaensis And Steroid

The heterogeneity of asthmatic diseases has plagued the decision-making of clinical treatment methods and caused considerable mortality due to its complicated mechanisms,which involves many autoimmune components,environmental factors,and epigenetic regulations.The series of studies in this thesis-researching studies have focused on the pathogenesis of multiple phenotypes of asthma and the potential multi-omics pharmacological mechanisms of traditional Chinese medicine Rhodiola wallichiana var.cholaensis(RWC).This series of studies have utilized high-throughput next-generation sequencing microbiome and highresolution metabolomics techniques and collected clinical samples from patients with multiple phenotypes of asthma(eosinophilic phenotypes,EA and non-eosinophilic phenotypes,NEA)for Multi-omics studies.By establishing the murine asthmatic models with different phenotypes,we performed an in-depth study on the pathogenesis of the asthmatic phenotypes.Besides,the immunological and the pharmacological mechanism of RWC treatment on the asthmatic phenotypes were also identified.At the same time,the content determination method of RWC injection and the potential fingerprint of RWC n-butanol extraction were also explored primarily.Many innovative research results have been obtained and are briefly described as follows:1.By studying the microbial flora in the induced sputum from asthmatic phenotypes,we found that the diversity,richness and evenness of airway microbiota in asthma patients decreased significantly,especially in NEA phenotypes.Further studies have discovered significant differences in the composition of the lower respiratory microbiota between the two phenotypes of asthma and between healthy individuals and asthmatic phenotypes.5 species were reported to be closely related to lung function(spirometry)and disease progression.Pathogens potentially associated with other genera also exhibited different patterns of association in different interaction networks.These findings further suggested that these potential pathogens may play different roles in the development of different disease phenotypes.2.Identification of all differential metabolites in peripheral sera of asthmatic patients with the phenotypes was performed with UPLC-MS/MS techniques under the multivariate analysis for clarifying the differences within all groups.As a result,we built 8 OPLS-DA models,7 of which showed reliably predictive features.Finally,we identified 18 different metabolites and 10 perturbed metabolic pathways based on the robust and reliable discriminant models.Glycerol-phospholipids,retinol and sphingolipid metabolism pathways,as the first top 3 significantly changed metabolic pathways was identified herein.3.The chromatographic method for quantitively determining the content of n-butanol extraction of RWC injection was first tested.In detail,the durability of this quantitative method was further examined by analyzing its characteristics such as stability,repeatability,intermediate precision,linearity and accuracy.Quantitation on salidroside was also finished.Then,asthmatic murine models were established horizontally.Therapeutic results showed that dexamethasone and dexamethasone co-treatment with RWC injection improve lung function,improve Th2/Th1 imbalance,regulate Th17 cell level,affect regulatory T cells and the levels of related cytokines(e.g.IL-13 and IL-17 etc.),which indicate mechanism of immune regulation.4.Microbiological and metabolomics methods were further utilized to identify the airway microbiota structure,metabolic pattern and local metabolic pattern of the asthmatic lungs.By treating with RWC and dexamethasone,it was found that the drug treatment with individual medicine and co-treatment with dexamethasone and RWC could significantly impact the composition of microbiota in NEA,especially for the 5 genera and 5 species.Besides,there is a significant increase in the diversity of the airway microbiota of the EA phenotype.The function of the airway microbiota has predicted that the microorganisms colonized in the airway may exert their functions with their own differential enzymes.A total of 34 differential metabolites were further identified by metabolomics studies.These 34 remarkably differential metabolites were widely distributed in different OPLS-DA discriminant models,and 8 differential metabolic pathways were also identified.Among them,there are 3 pathways associated with disease and treatment identified as the most significant(including primary bile acid biosynthesis,glycerophospholipid metabolism,linoleic acid metabolism,etc.).5.With joint multi-omics analysis,we found that colonization of airway microbiota may be involved in the hormone resistance and susceptibility characteristics of asthmatic diseases,and has a potential direct impact on individual's systemic or pulmonary local metabolic patterns.The pharmacological network based on the existing knowledge from present series studies macroscopically shows the immunological and Omicspharmacological mechanisms of RWC.