Electron paramagnetic resonance investigation of in vivo free radical formation and oxidative stress induced by 2,4-dichlorophenol in the freshwater fish Carassius auratus.
In the present study, electron paramagnetic resonance coupled with spin-trapping technique was used, with alpha-phenyl-N-tert-butylnitrone (PBN) as a spin-trapping agent, to investigate free radical generation in freshwater fish with acute 2,4-dichlorophenol (2,4-DCP) poisoning. The PBN-radical adducts were detected in fish liver samples following treatments of 2,4-DCP (0.025, 0.05, 0.5, 5, or 25 mg/kg) 24 h after intraperitoneal (i.p.) injection and 2,4-DCP (0.5 mg/kg) at 2, 4, 8, 24, or 72 h after i.p. injection in Carassius auratus. The hyperfine splitting constants for the PBN-radical adducts are aN = 13.7 G, aH = 1.8 G, and g = 2.0058, which is consistent with those of PBN/hydroxyl radical (*OH). The results indicate that the hydroxyl radical is probably produced during acute intoxication of 2,4-DCP. The relative similarity in the kinetics (from 2 to 72 h) of superoxide dismutase activity induction and *OH generation implies that the generation of *OH possibly depends on the superoxide anion (O2*-). Superoxide anion (O2*-) might be the precursor radical undergoing the Haber-Weiss reaction to form *OH. Possible pathways for radical chain reactions in the formation of the hydroxyl radical in vivo after 2,4-DCP administration are proposed. Other parameters with respect to antioxidant defense (e.g., superoxide dismutase and catalase) and oxidative damage (lipid peroxidation level) indicate that the fish were subjected to oxidative stress induced by 2,4-DCP and that the mechanisms of oxidative stress possibly involve the in vivo stimulation of hydroxyl radical formation.
Luo
Yi
Y
State Key Laboratory of Pollution Control and Resources Reuse, School of Environment, Nanjing University, Nanjing 210093, China.
Wang
Xiao-rong
XR
Shi
Hua-Hong
HH
Mao
Da-Qing
DQ
Sui
Yun-Xia
YX
Ji
Liang-Liang
LL
D000818:Animals; D000871:Anthelmintics / Q000633:toxicity; D000975:Antioxidants / Q000737:chemistry; D002374:Catalase / Q000378:metabolism; D002733:Chlorophenols / Q000737:chemistry / Q000633:toxicity*; D003497:Cyclic N-Oxides; D004305:Dose-Response Relationship, Drug; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods*; D004785:Environmental Pollutants; D016166:Free Radical Scavengers / Q000494:pharmacology; D005609:Free Radicals; D005618:Fresh Water; D006054:Goldfish; D017665:Hydroxyl Radical; D007700:Kinetics; D015227:Lipid Peroxidation; D008956:Models, Chemical; D009589:Nitrogen Oxides / Q000494:pharmacology; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D019097:Spin Trapping / Q000379:methods*; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000737:chemistry; D013997:Time Factors
D000871:Anthelmintics; D000975:Antioxidants; D002733:Chlorophenols; D003497:Cyclic N-Oxides; D004785:Environmental Pollutants; D016166:Free Radical Scavengers; D005609:Free Radicals; D009589:Nitrogen Oxides; D013481:Superoxides; D017665:Hydroxyl Radical; C029217:phenyl-N-tert-butylnitrone; D002374:Catalase; D013482:Superoxide Dismutase; C004762:2,4-dichlorophenol; D010100:Oxygen