Effects Of Different Drugs On Oxidant Stress Factors In The Expression Of Cigarrete Pretreated Alveolar Macrophage

Objective(1) To evaluate the role of dexamethasone(DEX),theophylline,ambroxol and erythromycin in the course of treatment for oxidant/antioxidant imbalance in COPD by observing the effect of the four drugs on the expression ofγ-GCS,TGF-β1 and c-fos mRNA and its protein in cigarreate smoke extract pretreated rat alveolar macrophage(AM).(2) To evaluate the role of TGF-β1 .in the course of oxidant/antioxidant imbalance in COPD by observing the effect of TGF-β1 on the expression ofγ-GCS,c-fos mRNA and its protein in cigarreate smoke extract pretreated rat AM.Methods(1) Acquired the AM of rat by BAL were randomly divided into six groups: control group, CSE group, DEX group, theophylline group, ambroxol group and erythromycin group. The last four groups were added DEX,theophylline,ambroxol and erythromycin respectivetly and made the final concentrations are 2.5μg/ml,20μg/ml,11μmol/ml and 5μg/ml respectivetly,two hours later,except control group ,others were added CSE,while control group added PBS. Cells were collected at 24,48 and 12 hours. Immunocytochemistry was used to examine the protein expression ofγ-GCS-HS,TGF-β1 and c-fos, respectivetly. Cells were collected at 18,36 and 6 hours , in situ hybridization staining was used to examine the mRNA expression ofγ-GCS-HS,TGF-β1and c-fos, respectivetly.(2) Acquired the AM of rat were randomly divided into three groups:control group, CSE group and TGF-β1 group,and made TGF-β1 group final concentration was 3ng/ml,two hours later,except control group ,others were added CSE,while control group added PBS. Cells were collected at 20 and 10 hours. Immunocytochemistry was used to examine the protein expression ofγ-GCS-HS and c-fos,respectivetly. The total RNA of alveolar macrophage was extracted respectively at 10 and 5 hours , reverse transcription polymerase chain reaction(RT-PCR)was used to examine the mRNA expression ofγ-GCS-HS and c-fos ,respectivetly.Result(1) The effect of cigarrete smoke on the expression ofγ-GCSh,TGF-β1 and c-fos mRNA and its protein: The expression ofγ-GCS(0.493±0.090,0.468±0.065),TGF-β1(0.412±0.017,0.280±0.018)and c-fos(1.369±0.150,1.121±0.124)mRNA and its protein of CSE group were increased significantly as compared to control group(0.206±0.006,...