Differential effects of cigarette smoke on oxidative stress and proinflammatory cytokine release in primary human airway epithelial cells and in a variety of transformed alveolar epithelial cells.
Differential effects of cigarette smoke on oxidative stress and proinflammatory cytokine release in primary human airway epithelial cells and in a variety of transformed alveolar epithelial cells.
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DOI：
10.1186/1465-9921-7-132
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发表时间：
2006-10-24
影响因子：
5.8
通讯作者：
Rahman, Irfan
中科院分区：
文献类型：
--
作者：
Kode, Aruna;Yang, Se-Ran;Rahman, Irfan
香烟烟雾介导的气道和肺泡上皮细胞的氧化应激和炎症反应是吸烟相关肺部疾病发病机制中的重要过程。以前，单个细胞系被用于评估香烟烟雾的氧化和促炎作用，结果令人混淆。在这项研究中，一组人类和啮齿动物转化的上皮细胞系被用来确定香烟烟雾提取物（CSE）对氧化应激标志物，细胞毒性和促炎细胞因子释放的影响，并与原代人小气道上皮细胞（SAEC）的影响进行了比较。用不同浓度的CSE（0.2-10%）处理原代人SAEC、转化人（A549、H1299、H441）和啮齿动物（小鼠MLE-15、大鼠L2）肺泡上皮细胞20 min至24 h。通过乳酸脱氢酶释放试验、台盼蓝排除法和吖啶橙子和溴化乙锭双重染色评估细胞毒性。通过酶循环测定法测定谷氨酰胺浓度，通过使用脂质过氧化测定试剂盒测定4-羟基-2-壬烯醛水平。通过ELISA测量促炎细胞因子（例如IL-8和IL-6）的水平。通过免疫细胞化学和免疫印迹法评估转录因子NF-κB的核转位。香烟烟雾提取物剂量依赖性地耗尽谷胱甘肽浓度，增加4-羟基-2-壬烯醛（4-HNE）水平，并导致转化细胞系以及SAEC坏死。转化的细胞系均未显示出响应于CSE的细胞因子的任何显著释放。CSE可诱导SAEC原代细胞释放IL-8和IL-6，并呈剂量依赖性，这与NF-κB核转位有关。这项研究表明，原代，但不是转化，肺上皮细胞是一个合适的模型来研究炎症机制，在响应香烟烟雾。
Cigarette smoke mediated oxidative stress and inflammatory events in the airway and alveolar epithelium are important processes in the pathogenesis of smoking related pulmonary diseases. Previously, individual cell lines were used to assess the oxidative and proinflammatory effects of cigarette smoke with confounding results. In this study, a panel of human and rodent transformed epithelial cell lines were used to determine the effects of cigarette smoke extract (CSE) on oxidative stress markers, cell toxicity and proinflammatory cytokine release and compared the effects with that of primary human small airway epithelial cells (SAEC). Primary human SAEC, transformed human (A549, H1299, H441), and rodent (murine MLE-15, rat L2) alveolar epithelial cells were treated with different concentrations of CSE (0.2–10%) ranging from 20 min to 24 hr. Cytotoxicity was assessed by lactate dehydrogenase release assay, trypan blue exclusion method and double staining with acridine orange and ethidium bromide. Glutathione concentration was measured by enzymatic recycling assay and 4-hydroxy-2-nonenal levels by using lipid peroxidation assay kit. The levels of proinflammatory cytokines (e.g. IL-8 and IL-6) were measured by ELISA. Nuclear translocation of the transcription factor, NF-κB was assessed by immunocytochemistry and immunoblotting. Cigarette smoke extract dose-dependently depleted glutathione concentration, increased 4-hydroxy-2-nonenal (4-HNE) levels, and caused necrosis in the transformed cell lines as well as in SAEC. None of the transformed cell lines showed any significant release of cytokines in response to CSE. CSE, however, induced IL-8 and IL-6 release in primary cell lines in a dose-dependent manner, which was associated with the nuclear translocation of NF-κB in SAEC. This study suggests that primary, but not transformed, lung epithelial cells are an appropriate model to study the inflammatory mechanisms in response to cigarette smoke.
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DOI：
10.1164/rccm.2110101
发表时间：
2002-08-15
影响因子：
24.7
作者：
Rahman, I;van Schadewijk, AAM;De Boer, WI
通讯作者：
De Boer, WI
DOI：
10.1016/s0006-291x(03)00224-9
发表时间：
2003-03-21
影响因子：
3.1
作者：
Rahman, I;Gilmour, PS;Aruoma, OI
通讯作者：
Aruoma, OI
DOI：
10.1165/rcmb.2004-0006oc
发表时间：
2004-12-01
影响因子：
6.4
作者：
Marwick, JA;Kirkham, PA;Rahman, I
通讯作者：
Rahman, I
影响因子：
--
作者：
Onoue, S;Endo, K;Kashimoto, K
通讯作者：
Kashimoto, K
DOI：
10.1111/j.1749-6632.1993.tb39148.x
发表时间：
1993-05-28
期刊：
ANNALS OF THE NEW YORK ACADEMY OF SCIENCES-SERIES
影响因子：
--
作者：
PRYOR, WA;STONE, K;PACKER, L
通讯作者：
PACKER, L