Protein carbonylation in human bronchial epithelial cells exposed to cigarette smoke extract
- 241 Downloads
Cigarette smoke is a well-established exogenous risk factor containing toxic reactive molecules able to induce oxidative stress, which in turn contributes to smoking-related diseases, including cardiovascular, pulmonary, and oral cavity diseases. We investigated the effects of cigarette smoke extract on human bronchial epithelial cells. Cells were exposed to various concentrations (2.5–5–10–20%) of cigarette smoke extract for 1, 3, and 24 h. Carbonylation was assessed by 2,4-dinitrophenylhydrazine using both immunocytochemical and Western immunoblotting assays. Cigarette smoke induced increasing protein carbonylation in a concentration-dependent manner. The main carbonylated proteins were identified by means of two-dimensional electrophoresis coupled to MALDI-TOF mass spectrometry analysis and database search (redox proteomics). We demonstrated that exposure of bronchial cells to cigarette smoke extract induces carbonylation of a large number of proteins distributed throughout the cell. Proteins undergoing carbonylation are involved in primary metabolic processes, such as protein and lipid metabolism and metabolite and energy production as well as in fundamental cellular processes, such as cell cycle and chromosome segregation, thus confirming that reactive carbonyl species contained in cigarette smoke markedly alter cell homeostasis and functions.
KeywordsCigarette smoke extract (CSE) 2D-electrophoresis Human bronchial epithelial cells (16-HBE cells) Matrix-assisted laser desorption/ionization (MALDI) Protein carbonylation
Keyhole limpet haemocyanin conjugate
High molecular weight
Matrix-assisted laser desorption/ionization time of flight
This research was supported by the “Piano di Sostegno alla Ricerca 2016—Linea 2” (Università degli Studi di Milano).
- Bondì ML, Ferraro M, Di Vincenzo S, Gerbino S, Cavallaro G, Giammona G, et al. Effects in cigarette smoke stimulated bronchial epithelial cells of a corticosteroid entrapped into nanostructured lipid carriers. J Nanobiotechnology. 2014;29:12–46.Google Scholar
- Chang D, Sha Q, Zhang X, Liu P, Rong S, Han T, et al. The evaluation of the oxidative stress parameters in patients with primary angle-closure glaucoma. PLoS One. 2011;6:4–9.Google Scholar
- Colombo G, Rusconi F, Rubino T, Cattaneo A, Martegani E, Parolaro D, et al. Transcriptomic and proteomic analyses of mouse cerebellum reveals alterations in RasGRF1 expression following in vivo chronic treatment with delta 9-tetrahydrocannabinol. J Mol Neurosci. 2009;37:111–22.CrossRefPubMedGoogle Scholar
- Kode A, Rajendrasozhan S, Caito S, Yang SR, Megson IL, Rahman I. Resveratrol induces glutathione synthesis by activation of Nrf2 and protects against cigarette smoke-mediated oxidative stress in human lung epithelial cells. Am J Phys Lung Cell Mol Phys. 2008;294:L478–88.Google Scholar
- Morris PB, Ference BA, Jahangir E, Feldman DN, Ryan JJ, Bahrami H, et al. Cardiovascular effects of exposure to cigarette smoke and electronic cigarettes: clinical perspectives from the Prevention of Cardiovascular Disease Section Leadership Council and Early Career Councils of the American College of Cardiology. J Am Coll Cardiol. 2015;66:1378–91.CrossRefPubMedGoogle Scholar
- Piao MJ, Ahn MJ, Kang KA, Ryu YS, Hyun YJ, Shilnikova K, et al. Particulate matter 2.5 damages skin cells by inducing oxidative stress, subcellular organelle dysfunction, and apoptosis. Arch Toxicol. 2018;92(6):2077–91. https://doi.org/10.1007/s00204-018-2197-9.CrossRefPubMedPubMedCentralGoogle Scholar
- Su Y, Cao W, Han Z, Block ER. Cigarette smoke extract inhibits angiogenesis of pulmonary artery endothelial cells: the role of calpain. Am J Phys Lung Cell Mol Phys. 2004;287:L794–800.Google Scholar
- van Rijt SH, Keller IE, John G, Kohse K, Yildirim AÖ, Eickelberg O, et al. Acute cigarette smoke exposure impairs proteasome function in the lung. Am J Phys Lung Cell Mol Phys. 2012;303:L814–23.Google Scholar