| Objective To explore the effccts of CSE on phagocytosis function of monocyte-derived macrophage in COPD patients and oxidative damage.Methods The peripheral blood mononuclear cells of32COPD patients and32healthy people were isolated. Adherent monocytes were collected from the cells, cultured in vitro and induced monocyte-derived macrophages(MDM).The MDM from COPD patients were divided into2groups:control group (group A), CSE treatment group (group B).The MDM from healthy controls were also divided into2groups:control group (group C), CSE treatment group (group D). Toxin function of CSE on MDM was measured by MTT. Flow cytometry combined with laser scanning confocal microscopylo scan the phagocytic activity of Escherichia coli (E. coli) labeled by FITC among groups.The contents of TAC,MDA and GSH-PX in the cell cultured supematants were tested by Spectrophotometer.Results (1) The longer time and higher concentration is used, the lower MDM survival rate.(2) Flow cytometry suggest that compared with MFI in group C (56.90±6.68), the MFI in group A (20.22±2.22) and group D (47.95±5.35) decreased significantly (P<0.01);compared with MFI in group A, the data in group B (7.65±0.70) decreased significantly (P<0.01). The decreased rang of MFI between group A and group B (61.92±4.01%) was significantly higher than group C and group D (15.23±9.16%)(P<0.01).(3) Using laser under confocal microscope conducted that MDM Phagocytosis of E.coli labeled by FITC (FITC-E.coli) of COPD patients was lower than healthy controls, and with CSE treatments, MDM Phagocytosis of all groups were reduced,and compared with the fluorescence ahsorbance values in group C (87.33±17.2), the fluorescence absorbance values in group A (51.43±5.85) and group D (69.72±5.97) decreased significantly (P<0.01);compared with the fluorescence absorbance values in group A, the data in group B (14.11±3.36) decreased significantly (P<0.01). The decreased rang of the fluorescence absorbance values between group A and group B (71.81±8.79%) was significantly higher than group C and group D (20.06±8.20%)(P<0.01).(4)The TAC and GSH-PX levels were lower in group A (4.06±0.23,47.10±4.13) U/ml than that in group C (5.14±0.29,88.40±2.31) U/ml (Pall<0.01), MDA level was significantly higher in group A(4.75±0.53)nmol/ml than that in group C (2.10±0.28)nmol/ml.TAC and GSH-PX levels (3.07±0.32,26.75±6.19)U/ml were significantly decreased (P all<0.01) and MDA level (7.72±0.85) nmol/ml was increased (P<0.01) in group B than that in group A.TAC and GSH-PX levels (4.47±0.36,72.30±5.14) U/ml were decreased(P all<0.01) and MDA level (2.95±0.56)nmol/ml was increased (P<0.01) in group D than that in group C.The decreased rang of TAC and GSH-PX levels between group A and group B (24.51±6.71%,43.25±12.03%) were significantly higher than group C and group D (13.05±5.13%,18.21±5.79%)(P all<0.01). The increased rang of MDA level between group A and group B (38.20±5.58%) was significantly higher than group C and group D (26.17.±1.76%)(P<0.05).(5) Under resting state,the postive correlations were respectively existed between MFI and TAC,GSH-PX (r=0.523or0.818, P<0.05or P<0.01).The MFI was negatively correlated to MDA (r=0.501, P<0.05). The TAC was postively correlated to GSH-PX (r=0.615, P<0.01). The MDA was postively correlated to TAC and GSH-PX (r=0.596or0.615, P all<0.05). After CSE treatment, the postive correlations were respectively existed between MFI and TAC,GSH-PX (r=0.704or0.716, P all<0.01). The MFI was negatively correlated to MDA (r=0.522, P<0.05). The TAC was postively correlated to GSH-PX (r=0.714, P<0.01). The MDA was postively correlated to TAC and GSH-PX (r=0.712or0.532, P<0.01or<0.05).Conclusion:As CSE concentration and role time increased, its effect on MDM increased.,with a significant dose and time-related effect. Compared with healthy people, photolytic function for FITC-E.coli of MDM in COPD patients decreased significantly, it might be demonstrated that the recurrent and acute exacerbations of COPD patients may result from declining phagocytic function of macrophage The phagocytosis of MDM in COPD patients were reduced by CSE treatment, and damage of photolytic function of MDM from COPD patients was stornger than those from healthy people. CSE could cause anoxidative damage in MDM, there were positive correlations between the MDM phagocytic function damages. Oxidative stress response might be the reason why phagocytic activity of macrophage from COPD patients were declined. |
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