Phagocytosis Function Of Macrophage In COPD Patients

Objective To explore the phagocytic function of macrophage in patients with chronic obstructive pulmonary disease (COPD).Methods From Oct.2011to Mar.2012,30patients of COPD and25healthy volunteers were selected. The PBMCs were isolated with Ficoll density gradient centrifugation from blood of both patients with COPD and healthy control groups;then the monocytes were collected with adherent method. After12-days cultivation in culture medium with Granulocyte-macrophage colony-stimulating factor, GM-CSF, cells are differentiated into mature MDM. According to the experiment requirements, cells were divided into healthy and COPD groups, and were both treated with sulforaphane, respectively. Using flow cytometry assay the MDM phagocytosis of both fluorescein-labeled Escherichia coll (fluorescein-E. coll) and pHrodoTM-labeled Staphylococcus aureus (pHrodoTM-S. aureus) were tested with or without intervention in different groups. Meanwhile the total number of colony generated from MDM phagocytosis of E. coll was calculated and compared by colony counting method.Results (1) The optimized concentration for phagocytosis of fluorescein-E. coll and pHrodoTM-S. aureus were determined to be0.04mg/ml and0.06mg/ml, respectively;6hours was the optimized time point.(2) The tested phagocytosis of fluorescein-E. coll and pHrodoTM-S. aureus were225.37±23.50and223.43±28.18respectively in healthy group while the correspondent values were153.39±18.34and155.04±22.67in COPD group, which decreased significantly(F=17.75,15.70, P all<0.01).(3) In COPD patients, with intervention of sulforaphene, MDM phagocytosis of fluorescein-E. coll and pHrodoTM-S. aureus were180.46±7.88and186.25±3.63, respectively, which is significantly increased(F=17.75,15.70, P all<0.01). While in healthy group, the correspondent phagocytic values were227.24±22.62and241.00±23.69, respectively which does not make a significant difference.(4) In both COPD and healthy groups, the MDM phagocytosis ability of fluorescein-E. coll and that of pHrodoTM-S. aureus are not significantly different with or without intervention.(5) The amount colonies generated from MDM phagocytosis was(62.57±2.58)in healthy group,(46.81±1.99) in COPD group, which was significantly decreased (F=30.90, P all<0.01). In COPD patients, with the sulforaphene intervention, the amount colonies generated from MDM phagocytosis of E. coli was (54.38±1.53), which is significantly increased compared with that before intervention (F=30.90,P<0.01). For healthy group, with the sulforaphene intervention, the amount colonies generated from MDM phagocytosis of E. coli was (64.03±2.09), which are not significantly different between conditions in absence or presence of sulforaphene intervention.Conclusion The acute exacerbations of COPD patients may result from decrease of phagocytic function. Sulforaphane can enhance the phagocytic function of macrophage in COPD patients.