Cigarette smoke, a complex mixture of over 7000 chemicals, contains many components capable of eliciting oxidative stress, which may induce smoking-related disorders, including oral cavity diseases. In this study, we investigated the effects of whole (mainstream) cigarette smoke on human gingival fibroblasts (HGFs). Cells were exposed to various puffs (0.5-12) of whole cigarette smoke and oxidative stress was assessed by 2',7'-dichlorofluorescein fluorescence. The extent of protein carbonylation was determined by use of 2,4-dinitrophenylhydrazine with both immunocytochemical and Western immunoblotting assays. Cigarette smoke-induced protein carbonylation exhibited a puff-dependent increase. The main carbonylated proteins were identified by means of two-dimensional electrophoresis and MALDI-TOF mass spectrometry (redox proteomics). We demonstrated that exposure of HGFs to cigarette smoke decreased cellular protein thiols and rapidly depleted intracellular glutathione (GSH), with a minimal increase in the intracellular levels of glutathione disulfide and S-glutathionylated proteins, as well as total glutathione levels. Mass spectrometric analyses showed that total GSH consumption is due to the export by the cells of GSH-acrolein and GSH-crotonaldehyde adducts. GSH depletion could be a mechanism for cigarette smoke-induced cytotoxicity and could be correlated with the reduced reparative and regenerative activity of gingival and periodontal tissues previously reported in smokers.
Oxidative damage in human gingival fibroblasts exposed to cigarette smoke / G. Colombo, I. Dalle Donne, M. Orioli, D. Giustarini, R. Rossi, M. Clerici, L. Regazzoni, G. Aldini, A. Milzani, D.A. Butterfield, N. Gagliano. - In: FREE RADICAL BIOLOGY & MEDICINE. - ISSN 0891-5849. - 52:9(2012), pp. 1584-1596.
|Titolo:||Oxidative damage in human gingival fibroblasts exposed to cigarette smoke|
|Parole Chiave:||cigarette smoke; redox proteomics; human gingival fibroblasts; protein carbonylation; protein thiols; GSH-alpha,beta-unsaturated aldehyde adducts; free radicals|
|Settore Scientifico Disciplinare:||Settore BIO/06 - Anatomia Comparata e Citologia|
Settore BIO/17 - Istologia
Settore CHIM/08 - Chimica Farmaceutica
|Data di pubblicazione:||2012|
|Digital Object Identifier (DOI):||http://dx.doi.org/10.1016/j.freeradbiomed.2012.02.030|
|Appare nelle tipologie:||01 - Articolo su periodico|