吸烟对老年慢性阻塞性肺疾病稳定期患者中性粒细胞还原型辅酶Ⅱ氧化酶的影响

Effect of smoking on NADPH oxidase in elderly patients with steady phase of chronic obstructive pulmonary disease

目的探讨吸烟对老年慢性阻塞性肺疾病（COPD）稳定期患者外周血中性粒细胞膜蛋白质及还原型辅酶Ⅱ（NADPH）氧化酶p47-^PHOX因子磷酸化表达的影响。方法选择134名受试者，分为4组：（1）吸烟COPD组39例;（2）吸烟无COPD组36例；（3）无吸烟COPD组31例；（4）无吸烟无COPD组28例。细胞色素C还原法测外周血中性粒细胞释放超氧阴离子（O2^-）的水平。Western blot检测外周血中性粒细胞膜蛋白质和NADPH氧化酶p47-^PHOX因子磷酸化表达，并分析O2^-的水平与其肺功能、血气之间关系。结果吸烟COPD组及吸烟无COPD组外周血中性粒膜细胞蛋白质和NADPH氧化酶p47-^PHOX因子磷酸化（呈黑色）明显强于无吸烟无COPD组（呈浅灰色）。吸烟COPD组中性粒细胞释放的O2^-水平与其第1秒用力呼气容积（FEV1）、FEV1占预计值百分比（％）、FEV1／用力肺活量、动脉血氧分压呈负相关（r分别为-0．898、-0．878、-0．874、-0．890，均为P〈50．01）。结论吸烟能激活NADPH氧化酶产生O2^-，并且吸烟能使NADPH氧化酶亚单位p47-^PHOX发生磷酸化。

Objective To investigate the effect of smoking on the phosphorylation levels of peripheral blood neutrophil membrane protein and of nicotinamide adenine dinucleotide phosphate （NADPH） oxidase p47-^PHOX factor in elderly chronic obstructive pulmonary disease （COPD） patients who smoking. Methods One hundred and thirty four subjects were enrolled in the study, and were divided into four groups., group 1： 39 smoking cases with stable COPD; group 2： 36 smoking cases without COPD; group 3： 31 non-smoking cases with stable COPD; group 4： 28 non-smoking cases without COPD group. The level of superoxide anion released by peripheral blood neutrophils was measured by cytochrome C reduction method. The phosphorylation levels of peripheral blood neutrophil membrane protein andnicotinamide adenine dinucleotide phosphate （NADPH）oxidase p47-^PHOX factor were assayed by Western blot. The relationship of pulmonary function and blood gas with superoxide anion level were analyzed. Results Phosphorylation levels of peripheral blood neutrophil membrane protein and NADPH oxidase p47-^PHOX factor were significant higher in group 1 and group 3 than in group 4 （all P〈0.05）. The superoxide anion released by peripheral blood neutrophils in group 1 was negatively correlated with FEV1 , FEV1 percentage of the predicted value, FEV1/FVC and PaO2（ r=-0.898,-0.878,-0.874,-0.890, all P〈0.01）. Conclusions Smoking can activate NADPH oxidase to produce superoxide anion, and make the important cytosolic component of NADPH oxidase p47-^PHOX to the phosphorylation state.