Banks M A, Porter D W, Martin W G, Castranova V
Division of Respiratory Disease Studies, National Institute for Occupational Safety and Health, Morgantown, West Virginia 26505.
Toxicol Appl Pharmacol. 1990 Aug;105(1):55-65. doi: 10.1016/0041-008x(90)90358-2.
Rat alveolar macrophages (AM) were isolated by pulmonary lavage, allowed to adhere to a tissue culture flask, and then exposed to 0.45 +/- 0.05 ppm ozone. After exposures ranging from 0 to 60 min, the medium was decanted and cells were harvested. Cells were assayed for oxidant damage and media analyzed for leakage of intracellular components. Increasing length of exposure to ozone resulted in a decreased number of adherent AM and decreased cell viability. Resting and zymosan-stimulated chemiluminescence increased immediately after ozone exposure and reached a maximum at 15-30 min, then declined to initial levels after 60 min of ozone exposure. Lipid peroxidation and leakage of protein and K+ ions increased with increasing length of exposure to ozone, while leakage of reduced and oxidized glutathione increased through 30 min, then declined (reduced) or leveled off (oxidized). Activity of the Na+/K+ ATPase decreased with time while intracellular taurine concentration exhibited an initial rise, peaked at 30 min, and then returned to the untreated level. Leakage of taurine into the medium increased with time of exposure, suggesting that exposure of AM to ozone results in a shift from bound to free intracellular taurine. These data indicate that in vitro exposure of AM to ozone results in a time-dependent alteration of cell function, membrane integrity, and viability.
通过肺灌洗分离大鼠肺泡巨噬细胞(AM),使其贴附于组织培养瓶,然后暴露于0.45±0.05 ppm的臭氧中。在0至60分钟的暴露后,倾去培养基并收获细胞。对细胞进行氧化损伤检测,并对培养基进行细胞内成分泄漏分析。臭氧暴露时间延长导致贴壁AM数量减少和细胞活力下降。静息和经酵母聚糖刺激的化学发光在臭氧暴露后立即增加,并在15 - 30分钟达到最大值,然后在臭氧暴露60分钟后降至初始水平。脂质过氧化以及蛋白质和K +离子的泄漏随着臭氧暴露时间的延长而增加,而还原型和氧化型谷胱甘肽的泄漏在30分钟内增加,然后下降(还原型)或趋于平稳(氧化型)。Na + / K + ATP酶的活性随时间下降,而细胞内牛磺酸浓度最初升高,在30分钟达到峰值,然后恢复到未处理水平。牛磺酸向培养基中的泄漏随暴露时间增加,表明AM暴露于臭氧会导致细胞内牛磺酸从结合型转变为游离型。这些数据表明,体外将AM暴露于臭氧会导致细胞功能、膜完整性和活力随时间发生改变。