摘要目的 探讨支气管哮喘(哮喘)与慢性阻塞性肺疾病(慢阻肺)及其重叠综合征患者外周血中性粒细胞极性化的差异.方法 根据2011年慢阻肺全球创议(GOLD)、全球哮喘防治创议(GINA)及2012哮喘-慢阻肺重叠综合征共识标准,人选2013年4月至2014年6月南方医院呼吸内科门诊受试者中健康不吸烟、健康吸烟、哮喘、慢阻肺及重叠综合征各20例.采用密度梯度离心法分离外周血中性粒细胞,通过Zigmond小室观察中性粒细胞在甲酰甲硫氨酰-亮氨酰-苯丙氨酰(fMLP)浓度梯度刺激下的自发极性化率、任意极性化率和定向极性化率.Western印迹检测各组外周血中性粒细胞的钙库操纵性钙内流(SOCE)的重要组分基质相互作用因子(STIM)1、2及Orai1蛋白表达情况.结果 哮喘组、慢阻肺组、重叠综合征组中性粒细胞自发极性化率显著高于健康不吸烟组、健康吸烟组[(25.05±4.06)％、(16.20 ±4.46)％、(29.43±5.53)％比(7.27±0.99)％、(7.06±3.12％),均P＜0.01],哮喘组、重叠综合征组中性粒细胞定向极性化率显著高于健康不吸烟组、健康吸烟组[(14.62 ±2.26)％、(8.00±1.75)％比(5.12±1.28)％、(5.24±1.34％),均P＜0.05],而慢阻肺组中性粒细胞定向极性化率[(2.45±0.54)％]显著低于健康不吸烟组、健康吸烟组(均P＜0.01).哮喘组、慢阻肺组、重叠综合征组中性粒细胞STIM1、2和Orai1蛋白表达灰度值均显著高于健康不吸烟组、健康吸烟组(STIM1:1.63 ±0.14、0.88±0.41、1.29±0.22比0.26±0.14、0.38 ±0.12;STIM2:0.52 ±0.19、0.22 ±0.13、0.24 ±0.10比0.05 ±0.03、0.10±0.06;Orai1:0.56 ±0.04、0.39±0.05、0.48±0.05比0.13 ±0.04、0.13 ±0.03)(均P＜0.01).结论 哮喘、慢阻肺及其重叠综合征患者外周血中性粒细胞的极性化存在明显异常和差异,且其SOCE的组分表达上调.

abstractsObjective To explore the polarization of migration dynamics of neutrophils isolated from patients with asthma,chronic obstructive pulmonary disease (COPD) and asthma-COPD overlap syndrome (ACOS) compared with healthy smoking and non-smoking controls.Methods Recruited volunteers were classified as healthy controls,healthy smokers,asthma,COPD and ACOS at Nanfang Hospital from April 2013 to June 2014 according to the Global Strategy for the Diagnosis,Management and Prevention of COPD 2011,Global Strategy for Asthma Management and Prevention 2011 and Consensus on Overlap Phenotype COPD-asthma in COPD 2012.Neutrophils were freshly isolated from whole blood with density gradient technique.The proportion of polarized cells with gradient concentration of formyl-Met-Leu-Phe (fMLP) in Zigmond chamber and vital component of Store Operated Calcium Entry (SOCE) (stromal interaction molecule (STIM)1,2 and Orai1) in neutrophils was detected by Western blot.Results Asthma,COPD and ACOS neutrophils demonstrated a higher spontaneous polarization rate versus healthy controls and healthy smokers ((25.05 ±4.06)％,(16.20 ±4.46)％,(29.43 ±5.53)％ vs (7.27 ±0.99)％,(7.06 ± 3.12)％,all P ＜ 0.01),asthma and ACOS neutrophils showed a higher directed polarization rate ((14.62 ±2.26)％,(8.00 ± 1.75)％,all P ＜0.05),but COPD had a relatively lower rate of directional polarization rate than healthy controls and healthy smokers ((2.45 ± 0.54) ％ vs (5.12 ± 1.28) ％,(5.24 ± 1.34)％,all P ＜0.01).The vital component of SOCE in neutrophils from asthma,COPD and ACOS were all up-regulated versus healthy controls and healthy smokers (STIM1:1.63 ± 0.14,0.88 ± 0.41,1.29±0.22 vs 0.26 ±0.14,0.38 ±0.12; STIM2:0.52 ±0.19,0.22 ±0.13,0.24 ±0.10 vs 0.05 ±0.03,0.10 ±0.06; Orail:0.56 ±0.04,0.39 ±0.05,0.48 ±0.05 vs 0.13 ±0.04,0.13 ±0.03) (all P ＜ 0.01).Conclusions Asthma,COPD and ACOS neutrophils are intrinsically different than counterparts from healthy control subjects and healthy smokers.And vital components of SOCE from patient neutrophils are intrinsically up-regulated.