Effects Of Inhaled Corticosteroids,Aminophylline And Other Medications On Improving Inflammatory Responses Of Asthma

Asthma is a common environmental-gene interaction that leads to chronic airway inflammation,characterized by various inflammatory cells infiltration around the airway,high secretion of mucus and airway remodeling,accompanied by a deterioration of lung function and reversible or even irreversible ventilatory dysfunction.Inhaled corticosteroids（ICS）,β₂-agonists,aminophylline,montelukast sodium and other medications are commonly used to treat asthma.Although they have a certain controlling effect on relieving clinical symptoms,improving lung function and airway inflammation,the specific characteristics and differences of their effect in controlling the degree of inflammation and lung function are not yet clear.The results of previous case-control study found that the levels of inflammatory cytokines in plasma,such as soluble CD14（s CD14）and complement factor D（CFD,also known as adipsin）,were significantly negatively correlated with the risk and severity of adult asthma and pulmonary function index FEV₁/FVC<75%.Therefore,this study first analyzed the relationship between ICS,long-actingβ₂receptor agonists（LABA）,aminophylline and the plasma levels of s CD14 and CFD in adult asthma patients,as well as the relationship with lung function through a population cross-sectional study.Further,through animal experiments,this study explored in depth the differences in improving asthma mice lung and systemic inflammation response among the four medications,including inhaled corticosteroids,short-actingβ₂receptor agonists（SABA）,aminophylline,and montelukast sodium,to clarify their antagonizing effects on asthma inflammation.The main purpose of this study is to provide new ideas and clues for more scientific and accurate clinical treatment of asthma.Part I:Associations of ICS,LABA,aminophylline with lung function,plasma cytokines expressions in adults.Objective:To explore the relationship between inhaled corticosteroids（ICS）,long-actingβ₂-agonists（LABA）,and aminophylline with lung function as well as plasma levels of s CD14 and adipsin in adult asthma.Methods:A total of 881 healthy controls and 585 adult asthma patients were included in the study.Their personal information,lifestyle,family history of asthma,and other information of each subject,as well as the clinical symptoms,frequency of attack,severity stage,medication treatment history of asthma patients were collected.The lung function（FEV₁,FVC,PEF,and FEV₁/FVC）and plasma cytokines（s CD14 and CFD）expression levels of each participant were measured.Asthma patients were divided into ICS/LABA group（n=197）,ICS/LABA/aminophylline group（n=335）,and aminophylline group（n=53）based on their medication history.The relationships of asthma medications with lung function parameters（FEV₁,FVC,and PEF）as well as plasma s CD14 and adipsin expressions were assessed by multiple linear regression models.Results:In the three treatment groups,the aminophylline group had the highest percentage of patients with intermittent attacks and without cough and dyspnea symptoms,while the ICS/LABA group had the highest proportion of patients with persistent attacks and severe dyspnea and expectoration symptoms（P<0.05）.After adjusting for confounding factors,the FVC,PEF levels of ICS/LABA group and ICS/LABA/aminophylline group were not significantly changed compared with healthy residents（P>0.05）,but the levels of anti-inflammatory cytokines s CD14 and CFD in this two groups were significantly decreased（P<0.05）,with the largest decrease of 0.48（95%CI:-0.57,-0.40）and 0.75（95%CI:-0.83,-0.68）natural logarithmic units in the ICS/LABA group among the three medications groups;Compared with healthy controls,the levels of FVC,FEV₁,and PEF in patients with asthma in the aminophylline group were significantly lower by 0.48 L,0.69 L,and1.57 L,respectively（P<0.05）,significantly lower than those in patients with asthma in the ICS/LABA group and the ICS/LABA/aminophylline group（P<0.05）,while the plasma levels of anti-inflammatory cytokines s CD14 and CFD decreased by 0.23（95%CI:-0.37 to-0.09）and 0.25（95%CI:-0.37,-0.12）natural logarithmic units,respectively,The decrease was significantly lower than the decrease of 0.48（95%CI:-0.57,-0.40）and 0.75（95%CI:-0.83,-0.68）natural logarithmic units in the ICS/LABA group and 0.34（95%CI:-0.11,-0.26）and 0.42（95%CI:0.48,-0.35）natural logarithmic units in the ICS/LABA/aminophylline group.When stratified by intermittent and persistent attacks,compared with healthy residents,there was no significant difference in FVC and PEF in the ICS/LABA group,while FEV₁decreased by-0.29L and-0.42L,respectively,with the smallest decrease among the three groups;At the same time,the plasma s CD14 and CFD expressions in the ICS/LABA group decreased by 0.35（95%CI:-0.46,-0.24）,0.67（95%CI:-0.77,-0.57）,and 0.61（95%CI:-0.72,-0.51）,0.84（95%CI:-0.94,-0.74）natural logarithmic units during intermittent and persistent attack,respectively,and the decrease was still the largest among the three medication groups.Conclusions:The lung function level of adult asthma patients treated with ICS/LABA or ICS/LABA/aminophylline was significantly higher than that of patients treated with aminophylline alone,while the plasma levels of anti-inflammatory factors CFD and s CD14 in patients treated with aminophylline alone were significantly higher than those in patients treated with ICS/LABA or ICS/LABA/aminophylline.It suggests that ICS/LABA therapy may be more effective than aminophylline in improving lung function,but aminophylline has a stronger effect in reducing inflammation in adults with asthma.Part II:The antagonistic effect of budesonide and terbutaline sulfate,amino phylline,and montelukast sodium on ovalbumin-induced inflammatory respo nses in mice.Objective:To investigate the effects of budesonide and terbutaline sulfate（ICS combined with SABA）,aminophylline,and montelukast sodium on the improvement of lung and systemic inflammatory cells in an ovalbumin（OVA）-induced mouse model of asthma,as well as their impact on the plasma levels of anti-inflammatory cytokines soluble CD14（s CD14）and complement factor D（CFD）.Methods:OVA was used to construct a mouse asthma model.A healthy control group,an OVA asthma group,a budesonide atomization treatment group,an aminophylline injection treatment group,and a montelukast sodium intragastric treatment group were established.The mice in OVA asthma group and medications group were sensitized by intraperitoneal injection of 100μg OVA and 2 mg aluminum adjuvant on day 1 and day 14,and mice were given an aerosolized challenge with 1%OVA on days 22-27.The medications groups were given nebulized budesonide suspension（0.5 mg budesonide and 5 mg salbutamol sulfate）,intraperitoneal injection of aminophylline injection（20 mg/kg）,and oral administration of montelukast sodium chewable tablets（30 mg/kg）before each OVA aerosolized challenge.The weight and lung organ coefficient of mice in each group were recorded.The pathological changes in the lung tissue of each group of mice were observed through H&E staining,and white blood cell counts and proportions in bronchoalveolar lavage fluid（BALF）and peripheral blood were detected.The proportion of CD3⁺T cells,B cells,and NK cells,as well as the proportion of CD4⁺T and CD8⁺T in CD3⁺T in peripheral blood lymphocytes were evaluated by flow cytometry.The expression levels of s CD14 and CFD in plasma were detected by ELISA kit.Results:Compared with the healthy control group,the mice in the OVA asthma group lost weight on the 27th day,their lung organ coefficients increased,their lung tissues showed significant inflammatory damage,a large number of inflammatory cells infiltrated around the airways,the proportion of eosinophils in BALF and peripheral blood significantly increased,the proportion of CD3⁺T,CD8⁺T,NK cells in peripheral blood lymphocytes significantly increased,and the proportion of CD4⁺T and B cells and the ratio of CD4⁺/CD8⁺and the expression levels of plasma s CD14 and CFD were significantly decreased（All P<0.05）.Compared with the OVA asthma group,the degree of pulmonary inflammatory damage and infiltration of inflammatory cells around the airways in the three treatment groups were significantly decreased（P<0.05）;In the budesonide group,the proportion of NK cells in peripheral blood lymphocytes and eosinophils in BALF were significantly decreased,while the proportion of B cells significantly increased（P<0.05）;In the aminophylline group,the proportion of eosinophils in peripheral blood and the proportion of CD3⁺T,CD8⁺T,and NK cells in lymphocytes decreased significantly,while the proportion of CD4⁺T,B,and the ratio of CD4⁺/CD8⁺increased significantly（P<0.05）;In the montelukast sodium group,the proportion of eosinophils in BALF and peripheral blood,and CD8⁺T,and NK cells in lymphocytes significantly decreased,while the proportion of CD4⁺T,the ratio of CD4⁺/CD8⁺,and the expression level of plasma s CD14 and CFD significantly increased to normal levels（P<0.05）.Conclusions:There are significant differences in the antagonistic effects on OVA-induced lung and peripheral blood inflammatory responses among the three conventional asthma medications of budesonide combined with terbutaline sulfate,aminophylline,and montelukast.Among them,budesonide combined with terbutaline sulfate combined with terbutaline sulfate have a better effect on improving lung inflammation;aminophylline has a good effect on improving systemic inflammation and disturbance of peripheral blood lymphocytes;montelukast has a better effect on improving the number and proportion imbalance of inflammatory cells in the lung and the whole body,as well as increasing the levels of anti-inflammatory cytokines s CD14and CFD in plasma.