Cigarette Smoke Condensate Causes a Decrease of the Gene Expression of Cu–Zn Superoxide Dismutase, Mn Superoxide Dismutase, Glutathione Peroxidase, Catalase, and Free Radical-Induced Cell Injury in SH-SY5Y Human Neuroblastoma Cells
Cigarette smoking condensate (CSC) contains oxidant compounds able to generate superoxide. The aim of the present study was to investigate the effect of the exposure to CSC on: (1) free radical production, (2) the gene expression of the antioxidant enzymes Cu–Zn superoxide dismutase (SOD1), Mn superoxide dismutase (SOD2), Glutathione Peroxidase (GPx), and catalase (CAT), and (3) cell survival in human neuroblastoma SH-SY5Y cells. The results showed that exposure (24 h) to different concentrations (10–150 μg/ml) of CSC caused a dose dependent cell injury that was coupled to the maximal increase of free radical production. These events were prevented by the addition to the incubation medium of the scavenger Vitamin E (50 μM). Furthermore, CSC exposure caused a reduction of the gene expression of the antioxidant enzymes SOD1, SOD2, GPx, and CAT that was counteracted by Vitamin E (50 μM). These results suggest that CSC exposure can induce a free radical overcharge that may be responsible for the inhibition of antioxidant enzymes expression and cell injury in SH-SY5Y human neuroblastoma cells. In fact the scavenger vitamin E can block both cell injury and inhibition of SOD1, SOD2, GPx, and CAT induced by CSC exposure.
Cigarette smoke condensate Antioxidant enzymes Vitamin E
This is a preview of subscription content, log in to check access.
We are indebted to Dr. Anna Pannaccione, Division of Pharmacology, Department of Neuroscience, Federico II University of Naples and dr. Floriana Flamma, BAT Italia, for their valuable help. This study was partially supported by the grant PRIN 2007 to prof. Paolo Mondola.
Amoroso S, Gioielli A, Castaldi M, Di Renzo GF, Annunziato L (1999) In the neuronal cell line SH-SY5Y, oxidative stress-induced free radical overproduction causes cell death without any participation of intracellular Ca2+ increase. Biochim Biophys Acta 1452:151–160CrossRefPubMedGoogle Scholar
Balansky R, Mircheva Z, Blagoeva P (1994) Modulation of the mutagenic activity of cigarette smoke, cigarette smoke condensate and benzo[a]pyrene in vitro and in vivo. Mutagenesis 9:107–112CrossRefPubMedGoogle Scholar
Chaudhari M, Jayaraj R, Santhosh SR, Rao PV (2009a) Oxidative damage and gene expression profile of antioxidant enzymes after T-2 toxin exposure in mice. J Biochem Mol Toxicol 23:212–221CrossRefPubMedGoogle Scholar
Chaudhari M, Jayaraj R, Bhaskar AS, Rao PV (2009b) Oxidative stress induction by T-2 toxin causes DNA damage and triggers apoptosis via caspase pathway in human cervical cancer cells. Toxicology 262(2):153–161CrossRefPubMedGoogle Scholar
Fukano Y, Oishi M, Chibana F, Numazawa S, Yoshida T (2006) Analysis of the expression of heme oxygenase-1 gene in human alveolar epithelial cells exposed to cigarette smoke condensate. J Toxicol Sci 31:99–109CrossRefPubMedGoogle Scholar
Mondola P, Annella T, Santillo M, Santangelo F (1996) Evidence for secretion of cytosolic CuZn superoxide dismutase by Hep G2 cells and human fibroblasts. Int J Biochem Cell Biol 28:677–681CrossRefPubMedGoogle Scholar
Mondola P, Ruggiero G, Serù R, Damiano S, Grimaldi S, Garbi C, Monda M, Greco D, Santillo M (2003) The Cu, Zn superoxide dismutase in neuroblastoma SK-N-BE cells is exported by a microvesicles dependent pathway. Brain Res Mol Brain Res 110:45–51CrossRefPubMedGoogle Scholar
Perocco P, Mazzullo M, Broccoli M, Rocchi P, Ferreri AM, Paolini M (2000) Inhibitory activity of vitamin E and alpha-naphthoflavone on beta-carotene-enhanced transformation of BALB/c 3T3 cells by benzo(a)pyrene and cigarette-smoke condensate. Mutat Res 465:151–158PubMedGoogle Scholar
Pryor WA, Hales BJ, Premovic PI, Church DF (1983) The radicals in cigarette tar: their nature and suggested physiological implications. Science 220:425–427CrossRefPubMedGoogle Scholar
Rodgman A, Green CR (2003) Toxic chemicals in cigarette mainstream smoke—hazard and hoopla. Beit Tabakforsch Int 20:481–539Google Scholar
Smith CJ, Hansch C (2000) The relative toxicity of compounds in mainstream cigarette smoke condensate. Food Chem Toxicol 38:637–646CrossRefPubMedGoogle Scholar
Stringer KA, Freed BM, Dunn JS, Sayers S, Gustafson DL, Flores SC (2004) Particulate phase cigarette smoke increases MnSOD, NQO1, and CINC-1 in rat lungs. Free Radic Biol Med 37:1527–1533CrossRefPubMedGoogle Scholar
Tsuda S, Matsusaka N, Ueno S, Susa N, Sasaki YF (2000) The influence of antioxidants on cigarette smoke-induced DNA single-strand breaks in mouse organs: a preliminary study with the alkaline single cell gel electrophoresis assay. Toxicol Sci 54:104–109CrossRefPubMedGoogle Scholar