| Objective:To studythe expression and change of Treg and Th17 in senile achronic obstructive pulmonary,disease explore the roles of Treg / 。Th17 imbalance in senile COPDMethods:The study was composed of two parts: animal experiment and clinical trial.1. Determine the age of rats by observing the color of fur rats, weight, organ coefficient ratio,function of various organs, the expression of Sirtuin-1. Sprague- Dawley(SD) rats were divided adult(6 month) group and aged(24month) group,each group randomly divided into control group,COPD model, RA-treated group,Cs A-treated group.The rat COPD model was established by intratracheal instillation of lipopolysaccharide and exposed to cigarette smoke daily. The lung function measurements were carried out, and the pathological changes were observed.The number of Treg cell and Th17 cell were detected by flow cytometry.Proteins expression of FOXP3,ROR in lung were Observed by western blot.Expression of IL-10 in lung were Observed by immunohistochemistry(IH)。Proteins expression of IL-10,IL-17 in bronchoalveolar lavage fluid(BLAF) and plasma were Observed by enzyme-linked immuno sorbent assay.2. Clinical peripheral lung tissue samples were obtained from patients undergoing resection for lung cancer.peripheral blood of patients with COPD and healthy people.Those patients were divided into four groups: adult controls,adult COPD group,senile controls group,senile COPD group.Proteins expression of FOXP3,ROR in lung were Observed by western blot.Expression of IL-10 in lung were Observed by immunohistochemistry(IH) 。Proteins expression of IL-10,IL-17 in plasma were Observed by enzyme-linked immuno sorbent assay.Results: 1.Animal experiments:(1)Determine the age of rats by observing the color of fur rats, weight, organ coefficient ratio,function of various organs, the expression of Sirtuin-1, We define the 24-month old rats are Senile rats.(2)The results of rats pulmonary function showed that percentage of forced expiratory volume in first 0. 3 second to forced vital capacity(FEV0.3/FVC%),forced expiratory volume in first0. 3 second(FEV0.3),and peak expiratory flow(PEF) were all decreased in COPD group compared with control group(P<)0.05 and which were all improved in RA-treated group compared with COPD model group( P<0.05).In contrast,It’s were all decreased in RA-treated group compared with COPD model group(P<0.05). Senile group decreased compared with adult group(P<0.05).Lung pathological changes in COPD model group conformed morphological character of COPD,pathological changes of rats lung tissue in RA-treated group reduced compared with COPD model group,however,pathological changes of rats lung tissue in Cs A-treated group increase compared with COPD model group.pathological of rats lung tissue in senile group behaved worse compared with adult group.(3)The number of Treg cells were decreased in COPD group compared with control group(P<0.05)and which were increased in RA-treated group compared with COPD model group( P<0.05).In contrast,its were decreased in RA-treated group compared with COPD model group( P<0.05). Senile group decreased compared with adult group( P<0.05).The number of Th17 cells were increased in COPD group compared with control group(P<0.05)and which were decreased in RA-treated group compared with COPD model group( P<0.05).In contrast,its were increased in RA-treated group compared with COPD model group( P<0.05). Senile group increased compared with adult group( P<0.05).(4)Proteins expression of FOXP3 in lung were decreased in COPD group compared with control group(P<0.05) and which were increased in RA-treated group compared with COPD model group( P<0.05).In contrast,its were decreased in RA-treated group compared with COPD model group( P<0.05). Senile group decreased compared with adult group( P<0.05).The Proteins expression of ROR were increased in COPD group compared with control group(P<0.05) and which were decreased in RA-treated group compared with COPD model group(P<0.05).In contrast,its were increased in RA-treated group compared with COPD model group(P<0.05). Senile group increased compared with adult group(P<0.05).(5)Proteins expression of IL-10 in bronchoalveolar lavage fluid(BLAF) and plasma were decreased in COPD group compared with control group(P<0.05)and which were increased in RA-treated group compared with COPD model group( P<0.05).In contrast,its were decreased in RA-treated group compared with COPD model group(P<0.05). Senile group decreased compared with adult group(P<0.05).Proteins expression of IL-17 were increased in COPD group compared with control group(P<0.05)and which were decreased in RA-treated group compared with COPD model group(P<0.05).In contrast,its were increased in RA-treated group compared with COPD model group(P<0.05). Senile group increased compared with adult group(P<0.05). 2.Clinical experiments:(1)Proteins expression of FOXP3 in lung were decreased in COPD group compared with control group(P<0.05)Senile group decreased compared with adult group(P<0.05).The Proteins expression of ROR were increased in COPD group compared with control group(P<0.05).Senile group increased compared with adult group( P<0.05).(2)Proteins expression of IL-10 in plasma were decreased in COPD group compared with control group(P<0.05)Senile group decreased compared with adult group(P<0.05).The proteins expression of IL-17 were increased in COPD group compared with control group(P<0.05).Senile group increased compared with adult group( P<0.05).(3)Linear correlation analysis showed proteins expression of IL-10 were positively correlated with FEV1(%)(P<0.05).proteins expression of IL-17 were negative correlated with FEV1(%)(P<0.05).Concl :usions 1.The 24-month old rats are Senile rats. 2.The expression of Treg,foxp3, IL-10 were reduced in COPD compared with control group. The expression of Th17,ROR, IL-17 were increased in COPD compared with control group. The expression of Treg and Th17 was imbalance.This imbalance of Treg and Th17 further eminent in senile COPD.3.The expression of IL-10 were positively correlated with FEV1.The expression of IL-17 were negative correlated with FEV1(%). |
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