The Effect Of Different Route Administration Of Corticosteroids On Ariway Microbiome In Aecopd Patients

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The characteristics of airway microbiome in acute exacerbations of COPDObjective To analyze and compare the diversity and the structural of airway microbiome with acute exacerbations and stable COPD.Methods A total of 19 patients with acute exacerbations of COPD were actue group and 19 patients with stable COPD were stable group.We collected the induced sputum samples from these patients.Total bacterial DNA was extracted from the sputum samples.The V3-V4 region of the bacterial 16 S rRNA gene was amplified.High throughput sequencing analysis was performed on the purified,pooled sample using an Illumina HiSeq 2500 platform.The OTUs cluster and species annotation was made with 97% sequences identity.Then we analyzed the relative abundance,alpha diversity and beta diversity between the acute and stable groups.Results 1.The Shannon index was lower in the acute group than in the stable group [(2.76±0.86)vs.(3.26±0.54),P=0.040].The Simpson index was higher in the acute group than in the stable group [(0.16±0.11)vs.(0.095±0.058),P=0.022].2.There were significant differences in the microbiome structure between the acute group and the stable group(P<0.05).At the phylum level,the Proteobacteria,Fusobacteria,Saccharibacteria,Chloroflexi and SR1_[Absconditabacteria] were siginifanctly higher in the acute group than in the stable group(P<0.05).The Cyanobacteria,Acidobacteria and Spirochaetae were siginifanctly lower in the acute group than in the stable group(P<0.05).At the class level,the Betaproteobacteria and Fusobacteriia were siginifanctly higher in the acute group than in the stable group(P<0.05).The Clostridia was much lower in the acute group than in the stable group(P<0.05).At the order level,the Neisseriales,Fusobacteriales,Pasteurellales,Clostridiales and Bacillales were siginifanctly higher in the acute group than in the stable group(P<0.05).At the family level,the Neisseriaceae,Pasteurellaceae,Lachnospiraceae,Fusobacteriaceae and Family_XI were siginifanctly higher in the acute group than in the stable group(P<0.05).The Porphyromonadaceae was much lower in the acute group than in the stable group(P<0.05).At the genus level,the Haemophilus,Neisseria,Alloprevotella and Fusobacterium were signifanctly higher in the acute group than in the stable group(P<0.05).The Porphyromonas were much lower in the acute group than in the stable group(P<0.05).Conclusions The microbiome diversity of COPD exacerbations decreased significantly.The microbiome diversity in COPD exacerbations was lower than in the stable COPD.There were differences in the the sputum microbiome structure at phylum,class,order,family and genus levels between the two groups.Patients with acute exacerbations showed an enrichment of common pathogens.It demonstrated that there was bacterial dysbiosis in COPD exacerbations patients.CHAPTER ? The characteristics of airway microbiome in eosinophilic COPD exacerbationsObjective This study aimed to investigate the characteristics of the airway microbiome in eosinophilic AECOPD patients before and after treatment.Methods Thirty-six acute exacerbations of COPD patients were divided into two groups according to the level of peripheral blood eosinophils.14 patients were eosinophilic group(peripheral blood eosinophils?300/ul)and 22 patients were noneosinophilic group(peripheral blood eosinophils<300/ul).All patients were treated with prednisolone acetate tablets 40 mg QD for five days.Other treatments were treated routinely according to COPD guidelines.We observed the clinical symptoms,lung function,the arterial blood gas and inflammatory mediums(IL-8,TNF-?)in the two groups before and after treatment.Induced sputum were collected before and after treatment for 1 day,5 days and 14 days.The induced sputum samples were analyzed by Illumina Hiseq 2500 platform throughput sequencing of 16 Sr DNA V3-V4 region.Sequenciong results were analyzed by Paired-end reads methods.The final quality reads were assembled into operational taxonomic units(OTUs)at the 97% similarity level.Then the disrtribution abundance and diversity of sputum in the two groups were obtained.Results 1.The diversity and the microbial composition was not significantly different between the eosinophilic and noneosinophilic groups before treatment.2.The clinical symptoms improved in the two groups after five days treatment.The pulmonary function and Pa O2 increased significantly in the two groups after five days treatment.The Pa CO2,hs-CRP,serum and sputum supernatant IL-8 and TNF-?decreased siginificantly in the two groups after five days treatment.The level of hs-CRP decreased siginificantly in the two groups after five days treatment.There were no significant differences between the two groups.3.The Shannon index was significantly increased in the two groups compared the baseline level after 5 days treatment.The Simpson index was in contrast to the Shannon index after 5 days treatment.4.The Shannon index was significantly increased in the two groups compared the baseline level and the Simpson index was in contrast to the Shannon index after 14 days treatment.The Chao1 and Ace indices were significantly increased in the Noneos group after 5 days and 14 days treatment,while it was contrast to the Eos group.5.There were several significantly different species after 14 days treatment.The different species in the eosinophilic group were Prevotella-7,Actinomycetales,Actinomycetaceae,Actinomyces and Actinomyces.The different species in the noneosinophilic group were Granulicatella,Granulicatella,Carnobacteriaceae,Rothia,microcococcales and microcococcacaea.Conclusions The diversity and the richness of the airway microbiome in the eosinophilic and noneosinophilic group were similar in the initial stage of treatment.The microbial diversity were increased signifanctly in the two groups.The richness of the airway microbiome decreased in the eosinophilic group after treatment,while it increased in the noneosinophilic group.CHAPTER ? Effects of different routes of adminstration and courses of glucocorticoids on airway microbiome and clinical efficacy in patients with COPD exacerbationsObjective First,to compare the impact of inhaled budesonide and prednisolone acetate tablets on the sputum microbiome and the clinical efficacy of AECOPD.Second,to compare the impact of 5 days and 10 days prednisolone acetate tablets use on the sputum microbiome and the clinical efficacy of AECOPD.Methods Forty-eight AECOPD patients were randomly divided into three group.A group(Budesonide group,15 cases)were treated with inhaled budesonide(2 mg tid);B group(prednisolone 5 days group,16 cases)were treated with Prednisolone acetate tablets 40 mg Qd for five days;C group(prednisolone 10 days group,17 cases)were treated with Prednisolone acetate tablets 40 mg Qd for then days.Observe the clinical symptoms,lung function,the arterial blood gas,inflammatory mediums(IL-8,TNF-?)and fasting blood glucose(FPG)in the three groups bdfore and after treatment.Induced sputum were collected before and after treatment for 1 day,5days,10 days and 30 days.Total bacterial DNA was extracted from the samples.The V3-V4 region of the bacterial 16 S r RNA gene was amplified with a common primer pair combined with adapter sequences and barcode sequences.High throughput sequencing analysis was performed on the purified,pooled sample using an Illumina Hi Seq 2500 platform.The OTUs cluster and species annotation was made with 97% sequences identity.Then the disrtribution abundance and diversity of sputum in the three groups were obtained.Results 1.The clinical symptoms improved in the three groups after five days treatment.The pulmonary function and Pa O2 increased significantly in the three groups after five days treatment(P < 0.05).The Pa CO2,hs-CRP serum and sputum supernatant IL-8 and TNF-?decreased siginificantly in the three groups after five days treatment(P < 0.05).There were no significant differences between the three groups(P > 0.05).There were no significant differences in the level of FPG in the three groups(P > 0.05).The Shannon index was significantly increased in the three groups compared the baseline level,[(2.82±0.24)vs.(2.51±0.18),P<0.05],[(2.84±0.23)vs.(2.73±0.31),P<0.05] and [(2.81±0.36)vs.(2.59±0.34),P < 0.05] respictively.There were no significant differences between the three groups(P > 0.05).The Simpson index was in contrast to the Shannon index.2.The clinical symptoms,pulmonary function,blood gas,hs-CRP,serum and sputum supernatant IL-8,TNF-?were siginificantly improved after ten days treatment in the three groups(P < 0.05).There were no significant differences between the three groups(P > 0.05).There were no significant differences in the level of FPG in the three groups(P > 0.05).The Shannon index was significantly increased in the three groups compared the baseline level(P < 0.05)and there were no significant differences between the three groups(P > 0.05).The Simpson index was in contrast to the Shannon index.3.One-way ANOVA analysis showed that there was statistical siginificantly difference in the level of FPG in the three groups after thirty days treatment.The level of FPG increased higher in the C group than in the A and B groups(P < 0.05).There were statistical siginificantly differences in the level of Shannon index and Simpson index in the three groups after thirty days treatment.The Shannon index increased significantly higher in the group A compared to the B and the C group(P < 0.05).The Simpson index was in contrast to the Shannon index.There were statistical siginificantly differences in the level of Chao1 and Ace index in the three groups after thirty days treatment.The Chao1 and Ace indices were lower in the group A than in the group B and group C.4.There was no significant different species in the three groups before treatment,after 5 days treatment and after 10 days treatment by Lef Se analysis.After 30 days treatment,the different species in group A were Gammaproteobacteria,Proteobacteria,Betaproteobacteriales,Neisseriaceae,Fusobacteriales,Fusobacteria,Neisseria,Alloprevotella,Clostridia,Pasteurellaceae and Haemophilus.While the different species in group B were Leptotrichia,Flavobacteriales,and Wessellaceae;the different species in group C were Firmicutes,Pseudomonadales,Granulicatella and Carnobacteriaceae.Conclusions Budesonide and prednisolone have the same effects on the clinical efficacy.Budesonide has little effect on the airway microbiome in AECOPD patients,while prednisolone has an impact on airway microbiome.Long term prednisolone has significantly higher effect on diversity of ariway microbiome in patients with AECOPD than short term prednisolone.