The Study Of Th17 Cell And Cytokine IL-17A By Animal Model In Asthma With Negative Emotions

Objective:Depression and anxiety are the most common negative emotions in asthma patients.Asthma patients with negative emotions have poor disease control and poor hormone therapy.How to identify and treat such patients is a clinical problem that needs to be solved urgently.In the onset of asthma,the over-activation of Th17 cells and the overexpression of its cytokine IL-17 A can cause the chemotaxis of neutrophilbased inflammatory cells,leading to airway inflammation;in the onset of depression,Th17 cells are over-activated and The overexpression of the cytokine IL-17 A will over-activate the hypothalamic-pituitary-adrenal(HPA)axis and changes in the expression of glucocorticoid receptors to aggravate central inflammation.Based on this,we established an animal model of asthma with negative emotions to clarify the phenotype of airway inflammation in the lung area in mice with asthma with negative emotions,the immunophenotypes in lung tissue,peripheral blood,and brain tissue,and the degree of HPA axis activation.Moreover,we hypothesize that blocking IL-17 A may better reduce airway inflammation and HPA axis activation.Finally,we also explored that IL-17 A monoclonal antibody may work by interfering with the expression of epigenetic molecules.Our research will provide a theoretical basis for exploring the therapeutic targets of refractory asthma.Materials and Methods:1.We established a mouse model of asthma by atomizing ovalbumin,a mouse model of depression and anxiety with chronic unpredictable stress,and a mouse model of asthma with negative emotion by atomizing ovalbumin combined with chronic unpredictable stress.model.Use sugar water preference test and open field test to detect mouse neurobehavior;Acetylcholine induces to detect mouse airway reactivity,alveolar lavage fluid cytokine level and number of inflammatory cells,lung tissue HE staining to observe the pathological changes of mouse lungs,PAS staining of lung tissue was used to observe mouse goblet cell proliferation and mucus secretion,and to explore the phenotype of airway inflammation in the lung area with asthma and negative emotions.Flow cytometry and q PCR were used to detect the expression levels of CD4+T lymphocyte subtype-related transcription factors and cytokines in lung tissue,peripheral blood,and brain tissue,and explore the lung area with negative emotions,the central nervous system,and the immune phenotype of the whole body.Finally,Elisa was used to detect the expression of HPA axis related factors in lung tissue,brain tissue and peripheral blood.2.On the basis of the successful establishment of animal models,a mouse model of asthma with negative emotions in which IL-17 A neutralizing antibody intervenes by intraperitoneal injection of IL-17 A was constructed.Use sugar water preference test and open field test to detect mouse neurobehavior;Acetylcholine induces to detect mouse airway reactivity,alveolar lavage fluid cytokine level and number of inflammatory cells,lung tissue HE staining to observe the pathological changes of mouse lungs,PAS staining of lung tissue was used to observe mouse goblet cell proliferation and mucus secretion,and to explore the effect of IL-17 A monoclonal antibody intervention on the phenotype of airway inflammation in the lung area of mice with asthma and negative emotions.Flow cytometry and q PCR were used to detect the expression levels of CD4+T lymphocyte subtype-related transcription factors and cytokines in lung tissue,peripheral blood,and brain tissue,and explore the effects of IL-17 A monoclonal antibody intervention on asthma with less negative emotions the influence of Th17 cells and related factors in mouse lung area,central nervous system and whole body.Finally,Elisa was used to detect the expression of HPA axis related factors in lung tissue,brain tissue and peripheral blood to explore the impact of IL-17 A monoclonal antibody intervention on the HPA axis.3.Detect the expression changes of epigenetic molecules PRMT5 and MBD2 by immunohistochemistry and q PCR and use correlation analysis to study the mechanism of IL-17 A monoclonal antibody intervention in the activation state of Th17 cells in mice with asthma and negative emotions.Results:1.We successfully induced mouse asthma model,mouse negative emotion model,mouse asthma with negative emotion model and corresponding neutralizing antibody intervention model.2.We compared with the control group,the mice in the asthma group and the asthma with negative emotion group have a lower sugar water preference index,high airway reactivity,an increase in the total number of cells and neutrophils in the alveolar lavage fluid,and inflammation infiltration of lung tissue Obviously,the content of inflammatory cytokines increased in alveolar lavage fluid,lung tissue,brain tissue,and plasma supernatant,the expression of Th17 cell-related factors increased,and the HPA axis was activated.3.After IL-17 A monoclonal antibody intervention,the glycemic preference index of mice with asthma and negative emotions increased,the number of airway inflammatory cells was significantly reduced,and the lung tissue inflammatory cell infiltration(H & E staining)and mucus hypersecretion(PAS staining)were significantly reduced.The activation of Th17 cells and related factors is weakened,and the activation of HPA axis is weakened.4.The expression of MBD2 and PRMT5 in the asthma group with negative emotions was significantly increased,and the range widened,which was positively correlated with the expression of IL-17 A.The methylation regulatory molecules MBD2 and PRMT5 were involved in regulating the immune phenotype.Conclusion:1.The method of OVA combined with CUMS can successfully construct a mouse model of asthma with negative emotions.2.The lungs of mice with asthma and negative emotions have an inflammatory phenotype dominated by neutrophils,and the degree of inflammation is higher than that of mice with simple asthma.3.Mice with asthma and negative emotions have activated both regional immune and system regions,with Th17 cell immune phenotypes.4.IL-17 A monoclonal antibody can inhibit the activation of immune and system areas in areas where asthma is accompanied by negative emotions and may be a potential therapeutic target.5.IL-17 A monoclonal antibody may regulate Th17 cells through the expression of epigenetic molecules PRMT5 and MBD2.