Formation of intracellular free radicals in guinea pig airway epithelium during in vitro exposure to ozone
Formation of intracellular free radicals in guinea pig airway epithelium during in vitro exposure to ozone
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DOI：
10.1006/taap.1996.8073
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发表时间：
1997-03-01
影响因子：
3.8
通讯作者：
Qu, QS
中科院分区：
文献类型：
--
作者：
Chen, LC;Qu, QS
在体内暴露于臭氧（O-3）已被证明会导致气道上皮损伤和脂质过氧化。多不饱和脂肪酸的氧化已被证明产生过氧化氢和醛类，活性氧物质（ROS）作为中间体。臭氧化的这些产物可能与其他生物有机分子发生反应，造成细胞损伤。为了评估活性氧的产生，将豚鼠气道上皮细胞的融合原代培养物在具有液-气界面的Costar膜上生长，并暴露于0.2、0.4和0.6 ppm O-3。使用荧光染料二氢罗丹明-123监测暴露期间细胞内ROS的浓度。细胞内的ROS浓度增加后立即开始的O-3曝光，并持续到曝光期结束（长达1小时）。随着O3-浓度的增加，细胞内ROS的浓度也随之增加。为了确定在O-3暴露过程中产生的ROS的种类，气道上皮细胞灌注与二甲基亚砜（DMSO），甲酸钠（羟基自由基清除剂），NaN 3（过氧化氢酶抑制剂），或二乙基二硫代氨基甲酸酯（DEDC，超氧化物歧化酶抑制剂）之前和暴露期间，荧光强度进行了连续监测。而DMSO和甲酸钠都降低了ROS的浓度，DEDC和NaN 3没有影响。我们的结论是，羟基自由基，而不是H2 O2或超氧阴离子产生后立即开始在豚鼠气道上皮细胞中的暴露浓度依赖性的方式的O-3曝光。(C)北京：科学出版社.
In vivo exposure to ozone (O-3) has been shown to cause airway epithelial damage and lipid peroxidation. The oxidation of polyunsaturated fatty acids has been shown to produce hydrogen peroxide and aldehydes with reactive oxygen species (ROS) as intermediates. These products of ozonation may react with other bioorganic molecules and cause cellular damage. To assess the production of ROS, confluent primary cultures of guinea pig airway epithelial cells were grown on Costar membrane with a liquid-air interface and exposed to 0.2, 0.4, and 0.6 ppm O-3. The concentrations of intracellular ROS during the exposure were monitored using the fluorescent dye dihydrorhodamine-123. The intracellular concentration of ROS increased immediately upon the commencement of the O-3 exposure and persisted until the end of the exposure period (up to 1 hr). The concentration of ROS increased with increasing O-3 concentration. To determine the species of ROS produced during O-3 exposure, airway epithelial cells were perfused with dimethyl sulfoxide (DMSO), sodium formate (hydroxyl radical scavengers), NaN3 (catalase inhibitor), or diethyl-dithio carbamate (DEDC, superoxide dismutase inhibitor) prior to and during the exposure period and the fluorescent intensity was monitored continuously. While both DMSO and sodium formate decreased the concentration of ROS, DEDC and NaN3 had no effect. We concluded that hydroxyl radicals instead of H2O2 or superoxide anions were produced immediately following the commencement of O-3 exposure in guinea pig airway epithelial cells in an exposure concentration-dependent fashion. (C) 1997 Academic Press.