The first application of a chemiluminescence probe, 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo[1,2-a]pyrazin-3-one (MCLA), for detecting O2- production, in vitro, from Kupffer cells stimulated by phorbol myristate acetate.
The purpose of this study was to investigate the generation of superoxide anion radical, O2-, by Kupffer cells in vivo in rat liver. Phorbol myristate acetate (PMA) was infused into perfused rat liver which had been continuously infused with 2-methyl-6-[p-methoxyphenyl]-3,7-dihydroimidazo- [1,2-a]pyrazin-3-one (MCLA, a highly sensitive luminescence reagent for O2-) and the MCLA luminescence from the liver surface was detected with a sensitive photon counter. Under identical conditions, but without MCLA infusion, PMA and nitro blue tetrazolium were infused to verify generation of O2- by formation of formazan deposition. Based on MCLA luminescence and formazan deposition, both dependent on the O2- reaction, we have concluded that liver Kupffer cells generate O2- in vivo in response to stimuli such as PMA. Further, the generation of O2- by Kupffer cells may be by the same mechanism as displayed by macrophages.
Uehara
K
K
Department of Anesthesiology and Reanimatology, Gunma University School of Medicine, Japan.
Maruyama
N
N
Huang
C K
CK
Nakano
M
M
D000818:Animals; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000378:metabolism; D007093:Imidazoles; D066298:In Vitro Techniques; D007728:Kupffer Cells / Q000187:drug effects* / Q000378:metabolism*; D008163:Luminescent Measurements; D008297:Male; D011719:Pyrazines; D051381:Rats; D017208:Rats, Wistar; D013481:Superoxides / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014969:Xanthine Oxidase / Q000378:metabolism
D007042:Hypoxanthines; D007093:Imidazoles; D011719:Pyrazines; D013481:Superoxides; C063412:2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo(1,2-alpha)pyrazin-3-one; D019271:Hypoxanthine; D014969:Xanthine Oxidase; D013755:Tetradecanoylphorbol Acetate