| ObjectiveTo observe T-lymphocyte subsets in peripheral blood and bronchoalveolar lavagefluid (BALF) and the phenotypical changes of dendritic cells (DCs) in chronic obstructivepulmonary disease (COPD) rats treated with Bu-Fei Yi-Shen granules, and to investigateits mechanism on regulating of dendritic cells.MethodsRats were randomly divided into six groups: control group (CG), model group(MG), Bu-Fei Yi-Shen high-dosage group (HDG), medium-dosage group (MDG),low-dosage group (LDG) and dexamethasone group (DG).12rats in each group. Thestable COPD rat model was duplicated by using the repeated smoke inhalations andbacterial infections. From the week9through20week, the rats were administratedwith normal saline in CG and MG, high-dosage Bu-Fei Yi-Shen in HDG, me-dium-dosage Bu-Fei Yi-Shen in MDG, Low-dosage Bu-Fei Yi-Shen in LDG and dex-amethasone in DG. Tidal volume (VT),50%tidal volume expiratory flow (EF50) andpeak expiratory flow (PEF) were measured by animal lung function measure system.Pulmonary morphology were observed by hematoxylin and eosin stain. T-lymphocytesubsets (CD3+, CD4+, CD8+, CD4+/CD8+) in peripheral blood and BALF, and dendriticcells surface protein (OX62, MHCII, CD80, CD86) in BALF and the lungs weremeasured by Flow Cytometry(FCM).Results1General statusFrom week4, rats in model group were emaciated, depressed, less active, lessfood or water. The symptoms were improved after treatment of Bu-Fei Yi-Shenhigh-dosage, medium-dosage, low-dosage and dexamethasone, especially in Bu-FeiYi-Shen high-dosage, medium-dosage groups. Body weight of rats in each group increased with age, but it in MG was lower thanCG from week14(P<0.05). Body weight of HDG, MDG, LDG and DG increasedquickly from week12, but there were no significant statistical difference among the groups(P>0.05).2Lung functionVT, EF50, PEF decreased significantly in MG compared with CG from the4thweek (P<0.05or P<0.01), while they increased significantly in HDG, MDG comparedwith MG from the16th week (P<0.05or P<0.01). EF50, PEF in DG was higher thanMG at the20th week (P<0.05).There was no significant differences in VT, EF50, PEFamong the four treatment groups during the20weeks period (P<0.05).3Lung pathologyAt the end of the20th week, lung histology impaired obviously in MG, and it wasimproved in Bu-Fei Yi-Shen high-dosage, medium-dosage, low-dosage and dexame-thasone.4T-lymphocyte subsets analysis in peripheral bloodCD4+, CD4+/CD8+were lower in MG than CG (P<0.01), CD8+was higher thanCG(P<0.01); CD4+, CD4+/CD8+were higher in HDG, MDG than MG(P<0.05orP<0.01), while CD8+was lower(P<0.01); CD8+was lower in DG than MG andLDG(P<0.05or P<0.01);CD4+/CD8+was higher in HDG, MDG than LDG(P<0.01),CD8+was lower(P<0.05or P<0.01); CD4+, CD4+/CD8+was higher in HDG, MDG thanDG(P<0.05or P<0.01). There was no significant differences in CD3+among thegroups(P<0.05).5T-lymphocyte subsets analysis in BALFCD4+/CD8+were significantly lower in MG than CG (P<0.01), CD8+was higherthan CG(P<0.01); CD8+was significantly lower in HDG, MDG than MG(P<0.05orP<0.01); CD8+was lower in DG than MG(P<0.05); CD4+/CD8+was higher in HDGthan MDG(P<0.05), CD8+was lower than LDG(P<0.05); There was no significantdifference in CD3+,CD4+,CD4+/CD8+in HDG, MDG, LDG or DG (P<0.05).6DCs phenotype in lung single cell suspensionOX62, OX62+MHCII+, OX62+CD80+, OX62+CD86+were higher in MG than CG(P<0.01), while they were lower in HDG, MDG than in MG, LDG and DG (P<0.05orP<0.01), OX62+MHCII+, OX62+CD86+were lower in LDG than MG (P<0.05orP<0.01). OX62, OX62+MHCII+, OX62+CD86+were lower in DG than MG (P<0.05orP<0.01). 7DCs phenotype in BALFOX62, OX62+MHCII+, OX62+CD80+, OX62+CD86+were higher in MG than CG(P<0.01); OX62, OX62+MHCII+, OX62+CD80+, OX62+CD86+in HDG, MDG waslower than MG(P<0.01);. OX62, OX62+CD80+, OX62+CD86+in DG was lower thanMG(P<0.01);. OX62+MHCII+, OX62+CD80+, OX62+CD86+in HDG, MDG was lowerthan LDG(P<0.05or P<0.01).8Correlation analysisIn single cell suspension，the level of OX62+was significantly negative correlatedwith CD4+, CD4+/CD8+in peripheral blood (r=-0.475,-0.598), and was significantlypositive correlated with CD8+in peripheral blood and CD3+, CD4+, CD8+in BALF(r=0.365,0.483,0.531,0.519); the level of OX62+MHCII+significantly negative cor-related with CD4+, CD4+/CD8+in peripheral blood (r=-0.533,-0.681), and the level ofOX62+MHCII+was significant with CD8+in peripheral blood and CD3+, CD4+, CD8+in BALF (r=0.400,0.619,0.572,0.749); the level of OX62+CD80+was significantlynegative correlated with CD4+, CD4+/CD8+in peripheral blood (r=-0.422,-0.576), andthe level of OX62+CD80+was significant with CD8+in peripheral blood and CD4+inBALF (r=0.421,0.418); the level of OX62+CD86+was significantly negative corre-lated with CD4+, CD4+/CD8+in peripheral blood (r=-0.404,-0.588), and the level ofOX62+CD86+was significant with CD8+in peripheral blood and CD3+, CD4+, CD8+inBALF (r=0.421,0.471,0.543,0.485). In broncho alveolar lavage fluid，the level ofOX62+was significant negative correlations with CD4+, CD4+/CD8+in peripheralblood (r=-0.478,-0.484); the level of OX62+MHCII+was significant negative correla-tions with CD8+in peripheral blood(r=0.349); the level of OX62+CD80+was signifi-cant negative correlations with CD4+/CD8+in peripheral blood (r=-0.610), and thelevel of OX62+CD80+was significant with CD8+in peripheral blood and BALF(r=0.706,0.659); the level of OX62+CD86+was significant negative correlations withCD4+/CD8+in peripheral blood (r=-0.509), and the level of OX62+CD86+was signifi-cant with CD8+in peripheral blood (r=0.503).Conclusion1There are beneficial effect of Bu-Fei Yi-Shen granule in the treatment of stableCOPD, which can significantly improve symptom, and lung function, reduce thepathological damage in the lungs and airway.2Bu-Fei Yi-Shen granule can significantly improve immune dysfunction in COPDrats, decrease CD8+level, inhibit surface proteins expression in DCs. The reduc- tion of migration and maturation of DCs in the lung tissue might be involved in themechanism of Bu-Fei Yi-Shen granule in the treatment of copd. |
PDF Full Text Request