Sex-related responses to oxidative stress in primary cultured hepatocytes of European flounder (Platichthys flesus L.).
Effects of oxidative stress induced by xenobiotic compounds were studied in primary cultures of isolated hepatocytes of immature European flounder (Platichthys flesus L.) of both sexes caught in a relatively unpolluted area of the German Bight (North Sea). Cells were exposed to oxidative stressors such as 100 microM hydrogen peroxide (H2O2), 100 microM benzo[a]pyrene (B[a]p) and 50 microM nitrofurantoin (N-(5-nitro-2-furfurylidene)-1-aminohydantoin; NF) for 2 and 24 h. Cell mortality was determined with the use of the fluorescent ethidium homodimer-1 and calcein. Oxidative stress response was assessed by quantitative analysis of (1) intracellular reactive oxygen species (ROS) formation with dihydrorhodamine 123, (2) lipid peroxidation on the basis of concentrations of lipid hydroperoxides and the lipid peroxidation products malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE) and (3) cellular total oxidant-scavenging capacity (TOSC) using the TOSC assay (Winston et al., 1998). An increase in ROS formation was detected as early as 2 h after exposure to H2O2, B[a]p and NF. After 24 h, stress responses were lower, except following exposure to NF. The pattern of responses differed with the different oxidative stressors. Lipid peroxidation and the capacity to scavenge ROS were increased significantly in both sexes only after exposure to H2O2, whereas B[a]p and NF provoked sex-dependent responses. B[a]p-induced lipid peroxidation and increase in scavenging capacity were observed only in hepatocytes of females, whereas NF initiated these responses only in cells of males. Sex differences in oxidative stress response only after exposure to pro-oxidants that require enzymatic activation infer the importance of biotransformation pathways in stress responses. Because of their sensitivity to oxidative stress, flounder hepatocytes provide a useful model for early risk assessment of xenobiotics.
Winzer
K
K
Biologische Anstalt Helgoland in der Stiftung Alfred Wegener Institut, Notkestrasse 85, 22607, Hamburg, Germany.
Winston
G W
GW
Becker
W
W
Van Noorden
C J
CJ
Köehler
A
A
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D005432:Flounder / Q000378:metabolism*; D015227:Lipid Peroxidation; D008099:Liver / Q000378:metabolism*; D008297:Male; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D012727:Sex Characteristics
D017382:Reactive Oxygen Species