The Influence Of Muscarinic Antagonism And Inhaled Corticosteroids On Clinic Effect And Airway Inflammation Of Different Types In Patients With COPD

[Objective]To observe the influence of muscarinic antagonism on clinic symptom,quality of life,pulmonary function test,airway inflammation of different types patients with chronic obstructive pulmonary disease(COPD),in order to find the theoretical basement for the anticholinergic bronchodilator treatment in patients with COPD.[Methods]1.103 acute COPD patients in exacerbation were enrolled in this study.All the subjects accepted the test of pulmonary function and bronchodilatation.Record clinic symptom scores,quality of life scores,FEV₁ and FEV₁ percent predicted.2.Sputum induction and analysisAll the subjects were premedicated with 200μg of inhaled salbutamol.Induction was performed using hypertonic saline(3%)aerosolised by compressed nebuliser for 15～20min.Entire sputum was collected in a plastic container.Select sputum and weigh sputum,then incubate with 4×volume 0.1%dithiothreitol(DTT)for 15min. Rock on bench rocker for 15 minutes,then mix with equal volume(to DTT)of phosphate buffered saline(PBS)and rock for 5 minutes.Filter through 48μm nylon gauze.Centrifuge at 2000r/min for 10 minutes.The supernatant was separated and stored at -70℃for future measurements the concentrations of LTB4,TNF-αand MMP-9.The cell pellet resuspended in 1ml PBS.Samples were discarded if squamous epithelial cells contamination was 20%or more.Total cell counts was determined in a haemocytometer.Make cell slides and Wright's staining.An overall differential cell counts on 400 nucleated non-squamous cells was performed by a examiner.3.All the patients were classified into two groups:tiotropium group, corticosteroids group.Then according to eosinophilic cell counts two groups were classified into Group A and Group B,Group A:bronchodilatation test was negative and sputum eosinophilic cell counts≥3%;Group B:neutrophil counts significant increasing,and sputum eosinophilic cell counts＜3%.The tiotropium groups were treated with tiotropium 18μg once a day;The corticosteroids groups were treated with ICS 250μg twice a day,recording clinic symptom scores,quality of life scores,and redetecting pulmonary function.After the treatment induced sputum was analysed and measured the concentrations of LTB4,TNF-α,MMP-9.[Results]1.There were no significant differences in common data including sex,age,smoking history and pulmonary function test in A and B groups at baseline(P＞0.05).2.The comparison of every maker between A and B groups in patients with COPD(1)Clinic symptom scores:Tiotropium group:there were no differences in clinic symptom scores in A and B groups before treatment(5.8±1.4)and(5.6±1.6)(P＞0.05).After treatment,A,B groups in clinic symptom scores decreased(4.0±1.2,4.2±1.0,respectively).There were significant difference between before treatment and after treatment(P＜0.05).ICS group:there were no differences in clinic symptom scores in A and B groups before treatment((5.2±1.6)and(5.3±1.5)(P＞0.05).After treatment,A,B groups in clinic symptom scores decreased(4.2±1.2,4.3±1.0,respectively).There were significant difference between before treatment and after treatment(P＜0.05)There were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).(2)Quality of life scores: Tiotropium group:there were no differences in quality of life scores in A and B groups before treatment(7.7±1.4 vs 8.0±1.3)(P＞0.05).After treatment,the patient groups on quality of life scores decreased(6.4±1.2,6.6±1.3,respectively). There were significant difference between before treatment and after treatment(P＜0.05).ICS group:there were no differences in quality of life scores in A and B groups before treatment(7.2±1.0 vs 7.8±1.3)(P＞0.05).After treatment,group A on quality of life scores decreased(6.2±1.2),there were significant difference between before treatment and after treatment(P＜0.05)But in group B,.there were no significant difference after treatment(P＞0.05).In group A:the quality of life scores for Tiotropium group decreased,but the quality of life scores for ICS group didn't decreased after treatment,there were significant difference for the two groups after treatment(P＜0.05).In group B:the quality of life scores decreased for Tiotropium group and ICS group after treatment,there were no significant difference for the two groups after treatment(P＞0.05).(3)FEV₁:Tiotropium group:increased from(1.08±0.22,1.20±0.24)L to(1.49±0.14, 1.45±0.18)L after treatment in the group A and B respectively.There were significant difference in FEV₁ between before treatment and after treatment(P＜0.05).ICS group:increased from(1.15±0.20,1.19±0.25)L to(1.45±0.23,1.42±0.21) L in the group A and B respectively.There were significant difference in FEV₁ between before treatment and after treatment(P＜0.05).There were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).(4)FEV₁%Tiotropium group::increased from(44.3±12.3,41.7±12.1)%to(53.3±13.8, 55.1±14.2)%after treatment in the group A and B respectively.There were significant difference in FEV₁ between before treatment and after treatment(P＜0.05).ICS group:increased from(42.3±11.1,44.1±12.1)%to(52.6±14.3, 55.1±14.6)%after treatment in the group A and B respectively.There were significant difference in FEV₁%between before treatment and after treatment(P＜0.05).There were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).(5)Sputum total and differential cell counts:Tiotropium group:after treatment:the percentage of neutrophils were declined from 75%to 40%(P＜0.05)in the groupB,but total cell counts and the percentage of eosinophils were unchanged after treatment;in the group A,total cell counts and the percentage of eosinophils were unchanged after treatment.ICS group:the percentage of eosinophils were declined from 7%to 3.0% after treatment in the group A,there were significant difference between before treatment and after treatment(P＜0.05).In the group B,total cell counts,the percentage of neutrophils and eosinophils did not differ after treatment.In group A:the percentage of eosinophils for Tiotropium group were not declined,but the percentage of eosinophils for ICS group were declined after treatment, there were significant difference for the two groups after treatment(P＜0.05).In group B:the percentage of neutrophils for Tiotropium group were declined,but the percentage of neutrophils for ICS group were not declined after treatment,there were significant difference for the two groups after treatment(P＜0.05).(6)LTB4:Tiotropium group:the sputum concentration of LTB4 in the group B was decrease from(557.4±120.5)pg/ml to(484.8±110.3)pg/ml after treatment(P＜0.05).ICS group:There were no significant difference between before treatment and after treatment in group A and B(P＞0.05).In group A:there were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).In group B:the LTB4 for Tiotropium group decreased,but LTB4 for ICS group didn't decrease after treatment,there were significant difference for the two groups after treatment(P＜0.05).(7)TNF-α:Tiotropium group:There were no significant difference between before treatment and after treatment in group A and B(P＞0.05).ICS group:There were no significant difference between before treatment and after treatment in group A and B(P＞0.05).There were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).(8)MMP-9:Tiotropium group:There were no significant difference between before treatment and after treatment in group A and B(P＞0.05).ICS group:There were no significant difference between treatment and after treatment in group A and B(P＞0.05).There were no significant difference between Tiotropium group and ICS group after treatment(P＞0.05).[Conclusion]1.One subdivision in the patients with COPD manifested neutrophilic airway inflammation,however the other subdivision showed eosinophilic inflammation.2.Inhaled tiotropium could improve clinic symptom,quality of life and pulmonary function;Inhaled corticosteroids could improve clinic symptom,quality of life and pulmonary function in the eosinophilic infiltation in patients with COPD. But in the neutrophilic infiltation in patients with COPD,it could improve pulmonary function;there were no significant difference in quality of life3.Inhaled tiotropium could reduce neutrophilic airway inflammation in the neutrophilic infiltation in patients with COPD,decrease the level of LTB4.Inhaled corticosteroids could reduce eosinophilic inflammation in the eosinophilic infiltation in patients with COPD.4 The eosinophil counts in induced sputum could be looked as a reference to deternmine whether inhaled tiotropium or corticosteroids for acute COPD patients in exacerbation.