The first application of a chemiluminescence probe, 2-methyl-6-[ P-methoxyphenyl]-3,7-dihydroimidazo[1,2-α]pyrazin-3-one (MCLA), for detecting O 2− production, in vitro, from Kupffer cells stimulated by phorbol myristate acetate

• Published in: FEBS letters 1993-12, Vol.335 (2), p.167-170
• Main Authors: Uehara, Kyoshi, Maruyama, Noboru, Huang, Chien-Kueng, Nakano, Minoru
• Format: Article
• Language: English
• Subjects: 2-Methyl-6-[ p-methoxyphenyl]-3,7-dihydroinudazo[1,2- a]pyrazin-3-one; Kupffer cell; Organ chemiluminescence; Phorbol myristate acetate superoxide
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(93)80722-7

The purpose of this study was to investigate the generation of superoxide anion radical, O − 2, 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 O 2 −) 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 O 2 −; by formation of formazan deposition. Based on MCLA luminescence and fonnazan deposition, both dependent on the O 2 −; reaction, we have concluded that liver Kupffer cells generate O 2 −; in vivo in response to stimuli such as PMA. Further, the generation of O 2 −; by Kupffer cells may be by the same mechanism as displayed by macrophages.