D'Anna Claudia, Cigna Diego, Costanzo Giorgia, Bruno Andreina, Ferraro Maria, Di Vincenzo Serena, Bianchi Laura, Bini Luca, Gjomarkaj Mark, Pace Elisabetta
Institute of Biomedicine and Molecular Immunology (IBIM), CNR, Palermo, Italy.
Mol Biosyst. 2015 Jun;11(6):1644-52. doi: 10.1039/c5mb00188a.
Smoking is strongly associated with diseases such as lung cancer and chronic obstructive pulmonary disease (COPD). Lung fibroblasts are crucial for the integrity of alveolar structure by producing extracellular matrix proteins which are required for attachment, structure, and function of alveolar epithelial cells. Despite the well-known association between cigarette smoke exposure and pulmonary and cardiovascular diseases, many questions remain regarding the mechanisms by which smoking induces diseases. The aim of this study is to detect differentially expressed proteins in human foetal lung cells (HFL-1) after 5 and 10% doses of cigarette smoke extract (CSE) exposure, combining two-dimensional electrophoresis (2DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In order to evaluate cellular ability to recover as well as lasting damage, we analysed the proteomic pattern 24 hours after the CSE removal (release). Eleven proteins had significant changes at various experimental points. Among these, 7 were up-regulated after CSE-treatments and 4 were down-regulated. Some spots seemed to be modified permanently or in a transient manner, in fact they returned to baseline levels after CSE-removal (normalisation after CSE release) and others were modified by selective CSE concentrations or only after release. MS identified, differentially expressed proteins are involved in stress response, mitochondrial activity, and aging. These findings may improve our understanding about molecular mechanisms underlying CSE caused damage and they may also integrate the comprehension of cigarette smoke effects on human health.
吸烟与肺癌和慢性阻塞性肺疾病（COPD）等疾病密切相关。肺成纤维细胞通过产生肺泡上皮细胞附着、结构和功能所需的细胞外基质蛋白，对肺泡结构的完整性至关重要。尽管吸烟与肺部和心血管疾病之间的关联已广为人知，但关于吸烟诱发疾病的机制仍存在许多问题。本研究的目的是结合二维电泳（2DE）和基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS），检测5%和10%剂量的香烟烟雾提取物（CSE）暴露后人胎儿肺细胞（HFL-1）中差异表达的蛋白质。为了评估细胞的恢复能力以及持续性损伤，我们在去除CSE（释放）24小时后分析了蛋白质组模式。在各个实验点，有11种蛋白质发生了显著变化。其中，7种在CSE处理后上调，4种下调。一些斑点似乎被永久或短暂修饰，实际上它们在去除CSE后（CSE释放后恢复正常）恢复到基线水平，而其他斑点则被选择性的CSE浓度修饰或仅在释放后被修饰。质谱鉴定出，差异表达的蛋白质参与应激反应、线粒体活性和衰老。这些发现可能会增进我们对CSE造成损伤的分子机制的理解，也可能会加深对香烟烟雾对人类健康影响的认识。
