Airway epithelial cell apoptosis and inflammation in COPD, smokers and nonsmokers
Airway epithelial cell apoptosis and inflammation in COPD, smokers and nonsmokers
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DOI：
10.1183/09031936.00063112
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发表时间：
2013-05-01
影响因子：
24.3
通讯作者：
Elborn, J. Stuart
中科院分区：
文献类型：
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作者：
Comer, David M.;Kidney, Joseph C.;Elborn, J. Stuart
为探讨慢性阻塞性肺疾病（COPD）患者的原代支气管上皮细胞（PBEC）对铜绿假单胞菌脂多糖（LPS）的反应与吸烟者和非吸烟者的PBEC不同，采用气液界面培养法对16例COPD患者、10例吸烟者和9例非吸烟者的PBEC进行了体外培养。在用铜绿假单胞菌LPS刺激之前，用CSE孵育培养物。ELISA法检测IL-6、IL-8的表达，荧光激活细胞分选仪检测Toll样受体（TLR）-4的表达。Western blotting和ELISA法检测核因子（NF）-κ B（B）的活化，Western blotting法检测MAPK和caspase-3的活性。采用Annexin-V染色和末端转移酶介导的dUTP缺口末端标记法评价细胞凋亡。COPD培养物中IL-8和IL-6的组成性释放最大。然而，CSE预处理后铜绿假单胞菌LPS刺激减少了COPD PBEC的IL-8释放，但增加了无气流阻塞的吸烟者和非吸烟者PBEC的IL-8释放。CSE治疗后，COPD培养细胞TLR-4表达、MAPK和NF-κ B活化降低，但在无气流阻塞的吸烟者或非吸烟者组中无此变化，这与细胞凋亡增加有关。CSE减弱COPD患者细胞对LPS的炎症反应，但对非吸烟者和无气流阻塞的吸烟者细胞无此变化。
We hypothesised that primary bronchial epithelial cells (PBECs) from subjects with chronic obstructive pulmonary disease (COPD) respond differently to Pseudomonas aeruginosa lipopolysaccharide (LPS) after cigarette smoke extract (CSE) exposure than PBECs obtained from smokers without airflow obstruction and nonsmokers.PBECs from 16 COPD subjects, 10 smokers without airflow obstruction and nine nonsmokers were cultured at air-liquid interface. Cultures were incubated with CSE prior to stimulation with P. aeruginosa LPS. Interleukin (IL)-6 and IL-8 were measured by ELISA and Toll-like receptor (TLR)-4 expression by fluorescence-activated cell sorter. Activation of nuclear factor (NF)-kappa B was determined by Western blotting and ELISA, and MAPK and caspase-3 activity by Western blotting. Apoptosis was evaluated using Annexin-V staining and the terminal transferase-mediated dUTP nick end-labelling methods.Constitutive release of IL-8 and IL-6 was greatest from the COPD cultures. However, CSE pretreatment followed by P. aeruginosa LPS stimulation reduced IL-8 release from COPD PBECs, but increased it from cells of smokers without airflow obstruction and nonsmokers. TLR-4 expression, MAPK and NF-kappa B activation in COPD cultures were reduced after CSE treatment, but not in the smokers without airflow obstruction or nonsmoker groups, which was associated with increased apoptosis.CSE attenuates inflammatory responses to LPS in cells from people with COPD but not those from nonsmoking individuals and those who smoke without airflow obstruction.