Pang Qi, Liu Xiaoju, Shi Kai, Zeng Xiaoli, Bao Hairong
Department of Gerontal Respiratory Medicine, First Hospital, Lanzhou University, Lanzhou 730000, China.
Department of Gerontal Respiratory Medicine, First Hospital, Lanzhou University, Lanzhou 730000, China. Email:
Zhonghua Yi Xue Za Zhi. 2014 Apr 1;94(12):895-8.
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).
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 non-CSE (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 thiobarbituric acid colorimetry and glutathione peroxidase (GSH-PX) by 5, 5'-dithiobis-2-nitrobenzoic acid (DTNB) method.
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 ± 0.5) vs (2.1 ± 0.4) µmol/L)]. And in COPD CSE and healthy CSE groups ((7.7 ± 0.9), (3.0 ± 0.6)µmol/L) were higher than those in COPD non-CSE and healthy non-CSE groups (all P < 0.01). At basic status, positive correlations existed between MFI and TAC, GSH-PX (r = 0.523, 0.818, P = 0.038, 0.001) while negative correlations between MFI and MDA (r = -0.501, P = 0.048) in COPD patients and after CSE treatment, the above relationships still existed (r = 0.704, 0.716, -0.522, P = 0.002, 0.002, 0.038).
Cigarette smoke can reduce the phagocytosizing ability of MDM in COPD patients. And it may be related with oxidative stress.
探讨香烟烟雾提取物（CSE）对慢性阻塞性肺疾病（COPD）患者单核细胞来源巨噬细胞（MDM）吞噬功能的影响。
2012年1月至2013年3月，从兰州大学第一医院32例稳定期COPD患者和32例健康对照者中分离外周血单核细胞。体外诱导并培养单核细胞成为MDM。将COPD患者和健康对照者的MDM分为COPD非CSE组（常规培养）、COPD CSE组（4% CSE处理6小时）、健康非CSE组（常规培养）和健康CSE组（4% CSE处理6小时）。采用流式细胞术（平均荧光强度，MFI）和激光扫描共聚焦显微镜（荧光灰度）检测MDM吞噬荧光素标记大肠杆菌（FITC - E.coli）的能力。采用邻菲罗啉比色法测定总抗氧化能力（TAC）。采用硫代巴比妥酸比色法测定丙二醛（MDA），采用5,5'-二硫代双（2-硝基苯甲酸）（DTNB）法测定谷胱甘肽过氧化物酶（GSH-PX）。
COPD非CSE组的MFI和荧光灰度（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）。且COPD CSE组（7.6±0.7，14.1±3.4）和健康CSE组（48.0±5.4，69.7±6.0）较COPD非CSE组和健康非CSE组下降更明显（均P<0.01）。COPD非CSE组的TAC和GSH-PX水平（（4.1±0.5），（47.1±4.1）U/ml）低于健康非CSE组（（5.1±0.6），（88.4±2.3）U/ml）。且COPD 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）分别低于COPD非CSE组和健康非CSE组（均P<0.01）。COPD非CSE组的MDA含量高于健康非CSE组[（4.8±0.5）对（2.1±0.4）μmol/L]。且COPD CSE组和健康CSE组（（7.7±0.9），（3.0±0.6）μmol/L）高于COPD非CSE组和健康非CSE组（均P<0.01）。在基础状态下，COPD患者中MFI与TAC、GSH-PX呈正相关（r = 0.523，0.818，P = 0.038，0.001），而MFI与MDA呈负相关（r = -0.501，P = 0.048），CSE处理后，上述关系仍然存在（r = 0.704，0.716，-0.522，P = 0.002，0.002，0.038）。
香烟烟雾可降低COPD患者MDM的吞噬能力。且可能与氧化应激有关。
