| Objective:To observe the difference of gut microbiota in children with asthma at different control levels.To investigate the effect of ketogenic diet on lung and gut microbiota in asthmatic mice.Methods:1.Children with asthma in our hospital from January 2020 to December 2020 were selected and divided into control group and uncontrolled group according to asthma control level.At the same time randomly selected in our hospital pediatric health examination of children as control group.After entering the group,general examination,lung function test,serum vitamin D test,gut microbiota 16S rRNA test,peripheral blood lymphocyte subsets level analysis and questionnaire survey were performed.The differences in the levels of each index among the three groups were compared,and the influencing factors of poor asthma control in children were analyzed.2.The OVA-Al(OH)3 combined induction and excitation was used to construct the asthma mouse model,and the mice were grouped and fed with ketogenic diet.The asthma situation of mice was evaluated by observing the behavioral state of mice in the excitation environment.hematoxylin-eosin(HE)staining was used to detect the pathological status of lung tissue;the feces of mice were detected by 16S rRNA sequencing to analyze the bacterial community structure and changes in the intestinal tract.The relative expression of HDACl mRNA was detected by real-time quantitative PCR.The content of short-chain fatty acids(SCFAs)in lung tissue was detected by liquid chromatography tandem mass spectrometry targeted metabolomics.Results:A total of 30 children with asthma were included.Control group 16 children;uncontrolled group 14 children;the control group of 12 children.There was no significant difference in lung function(FVC,FEV1,FEV1/FVC)among the three groups,and the value of FEF50 of lung function among the three groups was statistically significant(P<0.05).There were statistically significant differences in family allergy history,rhinitis,atopic dermatitis,food/drug allergy history and production mode among the three groups(P<0.05).The serum vitamin D level in the uncontrolled group was significantly lower than that in the control group(P<0.05).There was no significant difference in the diversity of gut microbiota among the three groups(P>0.05).At the genus level,Veillonella was the characteristic genus in the control group,and its abundance was significantly decreased in the uncontrolled group(P<0.05).ROC curve analysis suggested that Veillonella may have a certain value in judging the level of asthma control(specificity 28.6%,sensitivity 86.7%,area under ROC curve 0.82).In peripheral blood lymphocyte subsets,the differences in Thl and Th1/Th2 among the three groups were statistically significant(P<0.05).Further pairwise comparison,the control group compared with the control group,Th1,Th1/Th2 and killing T cell difference was statistically significant;compared with the control group,the difference of Thl and Th1/Th2 was still statistically significant(P<0.05).Non-targeted detection results of serum metabolism screened 79 differential metabolites,of which 27 differential metabolites can be enriched in the existing signaling pathways.3-hydroxybutyric acid may be the key metabolite in asthma control.2.The behavior of mice in ND group was more hyperactive and respiratory activity was more frequent than that in KD group.The average weight difference between the two groups during the experiment was statistically significant(P<0.05).HE staining results of lung tissue showed that there was a large amount of inflammatory cell infiltration in lung tissue of asthmatic mice in ND group.KD group had only a small amount of inflammatory cell infiltration,slightly less than ND group.There was no significant difference in the relative expression of HDAC1 mRNA and the content of short-chain fatty acids between the two groups(P>0.05).Beta diversity analysis of mouse feces 16s rRNA detection showed that there were significant differences in bacterial community diversity between the two groups.Conclusion:1.Vitamin D deficiency,allergic history,rhinitis,atopic dermatitis,history of food/drug allergy and production mode were all high risk factors affecting asthma attack.Traditional detection methods(lung function,vitamin D)have limited dynamic monitoring of asthma control level.Changes in gut microenvironment,such as changes in gut microbiota,have judgment value for improving control level and prognosis.2.Ketogenic diet can reduce some abnormal behaviors of asthmatic mice,reduce inflammatory cell infiltration and tracheal stenosis,and improve the imbalance of gut microbiota in asthmatic mice to a certain extent.Figure 25 Table 7 Reference 104... |
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