Increased oxidative stress in the RAW 264.7 macrophage cell line is partially mediated via the S-nitrosothiol-induced inhibition of glutathione reductase.
We investigated whether endogenously or exogenously produced nitric oxide (NO) can inhibit cellular glutathione reductase (GR) via the formation of S-nitrosothiols to decrease cellular glutathione (GSH) and increase oxidative stress in RAW 264.7 cells. The specificity of this inhibition was demonstrated by addition of a NO-synthase inhibitor, and met- or oxyhemoglobin. Using isolated GR we found that only certain NO donors inhibit this enzyme via S-nitrosothiol. Furthermore, we found that cellular GSH decrease is paralleled by an increase of superoxide anion production. Our results show that the GR enzyme is a potential target of S-nitrosothiols to decrease cellular GSH levels and to induce oxidative stress in macrophages.
Butzer
U
U
Department of General Surgery, University of Ulm, Germany.
Weidenbach
H
H
Gansauge
S
S
Gansauge
F
F
Beger
H G
HG
Nussler
A K
AK
D000818:Animals; D002460:Cell Line; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D005978:Glutathione / Q000031:analogs & derivatives / Q000378:metabolism / Q000494:pharmacology; D005980:Glutathione Reductase / Q000378:metabolism*; D008264:Macrophages / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008623:Mercaptoethanol; D051379:Mice; D009569:Nitric Oxide / Q000378:metabolism; D009603:Nitroso Compounds / Q000378:metabolism* / Q000494:pharmacology; D018384:Oxidative Stress / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D026422:S-Nitrosoglutathione; D026403:S-Nitrosothiols; D013481:Superoxides / Q000378:metabolism
D009603:Nitroso Compounds; D017382:Reactive Oxygen Species; D026403:S-Nitrosothiols; D013481:Superoxides; D009569:Nitric Oxide; D026422:S-Nitrosoglutathione; D008623:Mercaptoethanol; C032635:S-nitrosomercaptoethanol; C021315:S-nitrosocysteine; D005980:Glutathione Reductase; D005978:Glutathione; D003545:Cysteine