Short- and Long- Term Effects of Cigarette Smoke Exposure on Glutathione Homeostasis in Human Bronchial Epithelial Cells
Short- and Long- Term Effects of Cigarette Smoke Exposure on Glutathione Homeostasis in Human Bronchial Epithelial Cells
复制标题
DOI：
10.1159/000356633
复制
发表时间：
2013-01-01
影响因子：
--
通讯作者：
Garavaglia, Maria Lisa
中科院分区：
文献类型：
--
作者：
Bazzini, Claudia;Rossetti, Valeria;Garavaglia, Maria Lisa
背景资料：香烟烟雾提取物（CSE）是研究烟草烟雾在体内和体外作用的模型，它诱导细胞氧化应激并影响抗氧化谷胱甘肽系统。我们评估了CSE对气道上皮细胞的影响以及粘液药物S-羧甲基半胱氨酸赖氨酸盐（S-CMCLys）可能的细胞保护作用。研究方法：用荧光法检测人支气管上皮细胞（16-HBE）内还原型谷胱甘肽（GSH）和活性氧（ROS）水平，用RTPCR、Western blot和比色法检测GSH代谢途径相关酶的表达和活性。结果：CSE通过非凋亡依赖性途径，以时间和剂量依赖性方式显著增加细胞死亡率。短期（3小时）CSE暴露诱导ROS水平增加和GSH细胞内浓度下降。同时，谷胱甘肽过氧化物酶2和3，谷胱甘肽还原酶和谷氨酸-半胱氨酸连接酶的表达增加。S-CMC-Lys可有效地对抗这些作用。结论：CSE影响ROS水平、GSH浓度和GSH酶途径。这些作用可以在一定程度上被S-CMC-Lys逆转，其可以代表对抗CSE诱导的氧化细胞损伤的治疗工具。版权所有（C）2013 S. Karger AG，巴塞尔
Background: Cigarette smoke extract (CSE), a model for studying the effects of tobacco smoke in vivo and in vitro, induces cell oxidative stress and affects the antioxidative glutathione system. We evaluated the impact of CSE on airway epithelial cells and the possible cytoprotective effect of the mucolitic drug S-carboximethilcysteine lysine salt (S-CMCLys). Methods: Reduced glutathione (GSH) and reactive oxygen species (ROS) intracellular levels were evaluated by fluorimetry in human bronchial epithelial cells (16-HBE) and the expression and activity of enzymes of the GSH metabolic pathway were investigated by RTPCR, Western blot and colorimetric assays. Results: CSE significantly increased cell mortality in a time and dose dependent manner, via an apoptosis-independent pathway. Short-term (3 hours) CSE exposure induced an increase in ROS levels and a GSH intracellular concentration drop. In parallel, the expression of glutathione peroxidases 2 and 3, glutathione reductase and glutamate-cysteine-ligase was increased. S-CMC-Lys was effective in counteracting these effects. Conclusion: CSE affects ROS levels, GSH concentration and GSH enzymes pathway. These effects can be to some extent reversed by S-CMC-Lys, that could represent a therapeutic tool to counteract CSE induced oxidative cellular injuries. Copyright (C) 2013 S. Karger AG, Basel