Analysis Of Clinical And Metabonomic Characteristics Of Asthma With Different Pulmonary Function Types

Objectives1.Recruit asthma patients for standardized treatment for 3 months,and evaluate the clinical characteristics and drug treatment effects of asthma patients with different lung function types.2.Using metabonomics to study the intrinsic differences,disease mechanisms,and drug action pathways in patients with different lung functions,providing new biological targets for the diagnosis and treatment of diseases.Methods1.According to the inclusion and exclusion criteria,collect patients with chronic persistent moderate to severe asthma who were hospitalized in the Department of Respiratory and Critical Care Medicine of the People’s Hospital of Inner Mongolia Autonomous Region from November2018 to October 2022,complete a questionnaire survey,mainly including the patient’s basic personal information,clinical symptoms,disease history,laboratory tests,and related questionnaire evaluation.Retain blood and sputum samples of the patients,give Budesonide and Formoterol Fumarate Powder for Inhalation,Desloratadine Citrate Disodium Tablets,and Montelukast Sodium Chewable Tablets standard treatment for 3 months.The patient needs to undergo a follow-up visit every month,complete the corresponding questionnaire evaluation,and retain samples.According to the ratio of Forced expiratory volume in one second（FEV1）to Forced vital capacity（FVC）in the first second after bronchodilators,patients are divided into airflow restricted asthma（FEV1/FVC<0.7）and non airflow restricted asthma（FEV1/FVC≥0.7）.Fixed air flow obstruction（FAO+）is defined as the lung function of patients with both admission and exit airflow restrictions.The lung function of patients with both admission and exit airflow restrictions is defined as a Reversible airflow obstruction（FAO-）.The transition from admission to exit airflow restrictions is defined as an Inconsistent airflow obstruction（FAO±）.SPSS 26.0 software was used for data analysis.According to the nature of the data,statistical methods such as chi-square test,t-test,analysis of variance,or nonparametric test were used to analyze the data according to preset subgroups,and to compare the differences between groups of different lung function types and the treatment effect.2.Liquid chromatography mass spectrometer（LC-MS）was used to perform non targeted metabolomic analysis on the serum of patients in different groups.SIMCA-P 16.1 software was used to conduct multivariate data analysis.The variable weight values>1 and P<0.05 of the orthogonal partial least squares method for discriminating the analysis model were used as the screening criteria for differential metabolites,and the Pearson correlation coefficient was calculated through R script,To understand the correlation of significantly different metabolites,conduct metabolic pathway enrichment analysis on different metabolites based on the Kyoto Encyclopedia of genes and genomes（KEGG）database to identify significantly different metabolic pathways.Compare the metabolic differences between different groups before and after treatment horizontally,search for disease specific markers,analyze the effect of specific metabolic pathways on the disease,and compare the metabolic changes before and after treatment in different groups longitudinally to understand the differences in metabolic changes between different groups after drug action.Results1.Clinical data analysisComparing the clinical data at the time of admission（phase V0）,it was found that the levels of pulmonary ventilation function indicators FVC,FEV1,FEV1/FVC,and small airway function indicators such as peak expiratory flow（PEF）,maximum mid expiratory flow（MMEF）in patients in the FAO group were significantly higher than those in the other two groups,and there was no significant difference between FAO+and FAO±in the two comparisons,Patients in the FAO+and FAO±groups were more prone to wheezing than those in the FAO group（100%,85.7%,and 45%）.The family respiratory history of patients in the FAO+group was higher than those in the other two groups（90.9%,and 50%,and 42.9%）,with significant differences between the groups(P _wheezing=0.002;P _{family respiratory history}=0.035).There were no significant differences in other basic personal information,clinical symptoms,questionnaire evaluation,and basic laboratory tests.After 3 months of treatment（phase V3）,there were no significant differences in disease severity grading,control level grading,asthma control test（ACT）questionnaire,and asthma quality of life questionnaire（AQLQ）among the three groups of patients.In mental health assessment,the scores of self-rating anxiety scale（SAS）and self-rating depression scale（SDS）in the FAO group were significantly higher than those in the FAO+group,with significant differences between the groups(P_SAS=0.035,P_SDS=0.044).In terms of laboratory examination,the proportion of normal fractional concentration of exhaled nitric oxide（Fe NO）in the FAO-and FAO±groups was significantly higher than that in the FAO+group（42.1%,42.9%,vs.0%）,and the level of pulmonary function in the FAO+group was significantly lower than that in the FAO-and FAO±groups（P<0.05）.Longitudinal comparison of the V3 and V0 data of the three groups of patients showed that the proportions of patients in the FAO-,FAO±,and FAO+groups reaching intermittent status after treatment were 60%,50%,and 36.4%,respectively,and the proportions of patients achieving good control were 40%,21.4%,and 9.1%,respectively.The scores of ACT and AQLQ increased to varying degrees,with significant differences（P<0.05）.In mental health assessment,the score of SAS scale in the FAO±group decreased significantly after treatment（P<0.01）.In terms of laboratory examination,the value of Fe NO index in the FAO group significantly decreased compared to the baseline（P<0.05）;In the FAO±group,lung function indicators were significantly higher than baseline(P_FVC,FEV1,PEF<0.001,P_{FEV1/FVC,MMEF}<0.01);PEF in the FAO-and FAO+groups also significantly increased compared to baseline levels(P_PEF<0.01 in the FAO-group and P_PEF<0.05 in the FAO+group).Horizontal comparison of the difference in changes before and after treatment among the three groups was conducted to assess the difference in efficacy.The proportion of improvement in the severity rating questionnaire for the FAO-,FAO±,and FAO+groups was 80%,92.9%,and54.5%,respectively,and the proportion of improvement in the AQLQ questionnaire was 90%,85.7%,and 63.6%,respectively,but there was no significant difference between the groups（P>0.05）.In the mental health assessment,the SAS score of the FAO±group decreased by an average of 7 points,significantly higher than the decrease in the other two groups（P=0.013）,but there was no significant difference in the proportion of people who improved.In terms of laboratory examination,the proportion of Fe NO improvement in the FAO-,FAO±,and FAO+groups was 73.7%,69.2%,and 45.5%,respectively.However,there was no significant difference between the groups（P>0.05）.The improvement in pulmonary function in the FAO±group was significantly higher than that in the other two groups(P_FEV1,PEF<0.001,P_FVC,FEV1/FVC,MMEF<0.01).2.Metabolomic analysisThe number of metabolites identified in the positive ion mode of this project is 675,and the number of metabolites identified in the negative ion mode is 537,a total of 1212 metabolites.In the OPLS-DA score chart,each group of samples is separated on both sides of the y-axis,indicating that there are differences in metabolites between the groups.Through identification and screening,it was found that there were a total of 15 metabolites with significant differences between FAO±and FAO+before treatment.The most relevant metabolic pathways were mainly the carbohydrate digestion and absorption pathway involved in lactose and the galactose metabolic pathway.The energy metabolism level of the FAO+group was significantly higher than that of the FAO±group.There are a total of 10 metabolites with significant differences before and after treatment,and the most relevant metabolic pathway is the butyric acid metabolism pathway involved in malic acid.There are a total of 16 metabolites with significant differences before and after treatment in FAO±group,and a total of 21 metabolites with significant differences before and after treatment in FAO+group.There are four overlapping metabolites with significant differences between the two groups,namely,2-amino-1-phenylbutane,4-methylbenzyl alcohol,benzyl alcohol,and isopropanol.The above metabolites are mainly concentrated in the degradation pathway of xylene and aromatic compounds,and the level of sphingolipids in the FAO+group changes significantly before and after treatment,Among them,sphingomyelin（d18:1/18:0）is the most significantly involved in necrotic apoptosis.Conclusion1.Before treatment,it is difficult to distinguish between the FAO±and FAO+groups through existing clinical related questionnaires and examinations.The difference in metabonomics is mainly reflected in the significantly higher level of energy metabolism in the FAO+group than in the FAO±group,and it is expected that the level of energy metabolism can predict in advance the level of pulmonary function recovery in asthma patients with airflow restriction.2.Asthma patients with different lung function types have improved to varying degrees after 3months of treatment,with the largest improvement in the FAO±group and the smallest difference in changes before and after treatment due to airway remodeling in the FAO+group.The butyric acid metabolism pathway involved in malic acid plays a significant role in the treatment of the FAO group.The degradation pathway of xylene and aromatic compounds shared by FAO±and FAO+groups may be related to the mechanism of airway remodeling.The significant changes in sphingolipids levels before and after treatment in the FAO+group suggest that sphingolipids metabolism can be used as a specific therapeutic target for airway remodeling.