Use of fluorescent probes to assess the early sulfhydryl depletion and oxidative stress induced by mechlorethamine in human bronchial epithelial cells

Toxicology in Vitro : an International Journal Published in Association with BIBRA
S RappeneauFrancelyne Marano

Abstract

In this study, we report in vitro methods using fluorescent probes to assess thiol depletion and the oxidative stress induced by mechlorethamine (HN2), a nitrogen mustard, on a human bronchial epithelial cell line (16HBE14o-). Monobromobimane (mBBr) and 2',7'-dichlorofluorescin-diacetate (H(2)DCF-DA) were respectively used to monitor the intracellular thiol and peroxide levels. Fluorescent measurements were realized on gated live cells with a flow cytometer and a microspectrofluorimeter. Results clearly show that HN2 induced an early reduction of free sulfhydryl groups inside the cell correlated with an increase in the intracellular peroxides content. HN2 effects were time and dose dependent. The use of these fluorescent probes provide a useful and rapid methods for future screening of protective molecules against the early sulfydryl depletion and oxidative stress induced by HN2 on human airway epithelium.

References

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Citations

Jun 3, 2005·Metabolism: Clinical and Experimental·Nadjat OuslimaniJean-Louis Beaudeux
Dec 3, 2003·Toxicology and Applied Pharmacology·James F DillmanJohn J Schlager
Mar 23, 2013·Archiv der Pharmazie·Xianwen FangJia Zhou
Mar 4, 2015·Pediatric Blood & Cancer·Giovanni PaganoFederico V Pallardó
Jun 30, 2012·Critical Reviews in Toxicology·Feng Ru Tang, Weng Keong Loke
May 7, 2005·Free Radical Research·Emmanuelle Plantin-CarrenardFrançoise Braut-Boucher

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