Carnevali Stefano, Petruzzelli Stefano, Longoni Biancamaria, Vanacore Renato, Barale Roberto, Cipollini Monica, Scatena Fabrizio, Paggiaro Pierluigi, Celi Alessandro, Giuntini Carlo
Pulmonary Unit, Cardiothoracic Department, Pharmacology Unit, University of Pisa, 56124 Pisa, Italy.
Am J Physiol Lung Cell Mol Physiol. 2003 Jun;284(6):L955-63. doi: 10.1152/ajplung.00466.2001. Epub 2003 Jan 24.
Cigarette smoke is a mixture of chemicals having direct and/or indirect toxic effects on different lung cells. We investigated the effect of cigarette smoke on human lung fibroblasts (HFL-1) oxidation and apoptosis. Cells were exposed to various concentrations (1, 5, and 10%) of cigarette smoke extract (CSE) for 3 h, and oxidative stress and apoptosis were assessed by fluorescence-activated cell sorting and confocal laser fluorescence microscopy. Both oxidative stress and apoptosis exhibited a dose-response relationship with CSE concentrations. Lung fibroblasts also showed marked DNA fragmentation at the Comet assay after exposure to 10% CSE. Coincubation of HLF-1 cells with N-acetylcysteine (1 mM) during CSE exposure significantly reduced oxidative stress, apoptosis, and DNA fragmentation, whereas preincubation (3 h) with the glutathione-depleting agent buthionine sulfoximine (125 microM) produced a significant increase of oxidative stress. Cigarette smoke is a potent source of oxidative stress, DNA damage, and apoptosis for HFL-1 cells, and we speculate that this could contribute to the development of pulmonary emphysema in the lungs of smokers.
香烟烟雾是一种化学物质的混合物，对不同的肺细胞具有直接和/或间接的毒性作用。我们研究了香烟烟雾对人肺成纤维细胞（HFL-1）氧化和凋亡的影响。将细胞暴露于不同浓度（1%、5%和10%）的香烟烟雾提取物（CSE）中3小时，并通过荧光激活细胞分选和共聚焦激光荧光显微镜评估氧化应激和凋亡。氧化应激和凋亡均与CSE浓度呈剂量反应关系。在暴露于10% CSE后，肺成纤维细胞在彗星试验中也显示出明显的DNA片段化。在CSE暴露期间，将HLF-1细胞与N-乙酰半胱氨酸（1 mM）共同孵育可显著降低氧化应激、凋亡和DNA片段化，而用谷胱甘肽消耗剂丁硫氨酸亚砜胺（125 microM）预孵育（3小时）则会显著增加氧化应激。香烟烟雾是HFL-1细胞氧化应激、DNA损伤和凋亡的强大来源，我们推测这可能有助于吸烟者肺部肺气肿的发展。
