Induction of oxidative stress in murine cell lines by 3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX).
3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone (MX), the potent bacterial mutagen produced during chlorination of drinking water, was tested for the induction of oxidative stress in two murine cell lines: NIH 3T3 (fibroblasts) and L929 (fibrosarcoma cells). Following 1 h MX treatment at concentrations between 100 and 1000 microM, cellular stress conditions were monitored by measuring reactive oxygen species formation (ROS) and reduced glutathione levels (GSH). The kinetics of ROS formation and GSH depletion was investigated from 10 min to 1 h. MX caused detachment of cells at 1000 microM in L929 cells and at 300 microM in NIH 3T3 cells but the viability of the cells, measured by the trypan blue assay, decreased only by 20 and 7%, respectively, in 1h. MX increased ROS production in L929 cells in a dose-dependent manner, by 120% at 500 microM of MX in 1 h. The maximum ROS production was attained already in 10min. In NIH 3T3 cells, the ROS production was slightly, but not statistically significantly stimulated at 200 microM between 20 and 60 min. Concomitantly, MX decreased the intracellular content of GSH dose-dependently in both cell lines, by 48% in L929 cells at 500 microM of MX and 32% in NIH 3T3 cells at 200 microM of MX in one hour. The majority of this GSH depletion had occurred in 10 min. These findings indicate that MX induces oxidative stress in mammalian cells in vitro though the sensitivity of cells may differ for this effect.
Zeni
O
O
Interuniversity Centre for Interaction Between Electromagnetic Fields and Biosystems (ICEmB) at CNR-IREA, Via Diocleziano, 328-80124, Naples, Italy.
Salvemini
F
F
Di Pietro
R
R
Buonincontri
D
D
Komulainen
H
H
Romanò
M
M
Scarfí
M R
MR
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005354:Fibrosarcoma / Q000188:drug therapy / Q000378:metabolism; D005663:Furans / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism; D051379:Mice; D009153:Mutagens / Q000633:toxicity*; D041681:NIH 3T3 Cells / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000032:analysis
D005663:Furans; D009153:Mutagens; D017382:Reactive Oxygen Species; C054121:3-chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone; D005978:Glutathione