目的 探讨香烟烟雾提取物(CSE)对慢性阻塞性肺疾病(简称慢阻肺)患者单核细胞源性巨噬细胞(MDM)吞噬功能的影响.方法 选择2012年1月至2013年3月兰州大学第一医院慢阻肺稳定期患者32例,选择同期健康体检者32名,分离外周血单核细胞,体外诱导培养成MDM.分别将慢阻肺患者和健康体检者的MDM分为慢阻肺非CSE组(常规培养)、慢阻肺CSE组(4% CSE干预6h)、健康非CSE组(常规培养)、健康CSE组(4% CSE干预6h).以流式细胞术的平均荧光强度和激光共聚焦显微镜的荧光灰度值判断MDM吞噬荧光标记大肠杆菌(FITC-E.coli的能力,菲罗啉比色法检测细胞上清液总抗氧化能力(TAC),硫代巴比妥酸比色法检测丙二醛含量,改良Hafeman直接测定法检测谷胱甘肽过氧化物酶(GSH-PX)活力.结果 平均荧光强度、荧光灰度值:慢阻肺非CSE组(20.2±2.2、51.5±5.8)显著低于健康非CSE组(56.9±6.7、87.3 ±7.3),慢阻肺CSE组(7.6±0.7、14.1 ±3.4)和健康CSE组(48.0±5.4、69.7±6.0)分别显著低于慢阻肺非CSE组和健康非CSE组(均P<0.01).TAC、GSH-PX水平:慢阻肺非CSE组[(4.1±0.5)、(47.1±4.1)U/ml]显著低于健康非CSE组[(5.1±0.6)、(88.4 ±2.3)U/ml],慢阻肺CSE组和健康CSE组[(3.1±0.4)、(26.8±6.2) U/ml和(4.5±0.4)、(72.3±5.1) U/ml]分别显著低于慢阻肺非CSE组和健康非CSE组(均P<0.01).丙二醛水平:慢阻肺非CSE组[(4.8 ±0.5)μmol/L]显著高于健康非CSE组[(2.1±0.4)μmol/L],慢阻肺CSE组和健康CSE组[(7.7±0.9)和(3.0±0.6) μmol/L]分别显著高于慢阻肺非CSE组和健康非CSE组(均P<0.01).慢阻肺组平均荧光强度在基础状态下与TAC、GSH-PX水平呈正相关(r =0.523、0.818,P=0.038、0.001),与丙二醛水平呈负相关(r=-0.501,P=0.048),CSE干预后上述相关关系依然存在(r=0.704、0.716、-0.522,P=0.002、0.002、0.038).结论 香烟
Objective To explore the effects of cigarette smoke extract (CSE) on phagocytosizing function of monocyte-derived macrophages (MDMs) in patients with chronic obstructive pulmonary disease (COPD).Methods From January 2012 to March 2013,peripheral blood monocytes were isolated from 32 stable COPD patients and 32 healthy controls at First Hospital,Lanzhou University.MDM was induced and cultured from monocytes in vitro.The MDMs from COPD patients and healthy controls were divided into 4 groups of COPD non-CSE (conventional culture),COPD CSE (4% CSE treatment for 6 h),healthy nonCSE (conventional culture) and healthy CSE (4% CSE treatment for 6 h).Flow cytometry (mean fluorescence intensity,MFI) and laser scanning confocal microscopy (fluorescence grey level) were applied to detect the ability of MDM phagocytosed fluorescein-labeled Escherichia coli (FITC-E.coli).Total antioxidative capacity (TAC) was measured by o-phenanthroline colorimetry.Malondialdehyde (MDA) was measured by thiobarbituricacid colorimetry and glutathione peroxidase (GSH-PX) by 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) method.Results MFI and fluorescence grey level in COPD non-CSE group (20.2 ± 2.2,51.5 ± 5.8) significantly decreased than those in healthy non-CSE group (56.9 ± 6.7,87.3 ±7.3).And in COPD CSE (7.6 ±0.7,14.1 ±3.4) and healthy CSE groups (48.0 ±5.4,69.7 ±6.0) decreased more than those in COPD non-CSE and healthy non-CSE groups (all P 〈 0.01).The levels of TAC and GSH-PX in COPD non-CSE group ((4.1 ±0.5),(47.1 ±4.1) U/ml) were lower than those in healthy non-CSE group ((5.1 ± 0.6),(88.4 ± 2.3) U/ml).And in COPD CSE and healthy CSE groups ((3.1 ± 0.4),(26.8 ± 6.2) U/ml) and (4.5 ± 0.4),(72.3 ± 5.1) U/ml) were respectively lower than those in COPD non-CSE and healthy non-CSE groups (all P 〈 0.01).The content of MDA in COPD non-CSE group was higher than that in healthy non-CSE group [(4.8