Aflatoxin B(1) inhibits CD14-mediated nitric oxide production in murine peritoneal macrophages.
Aflatoxin B(1) (AFB(1)), a potent hepatocarcinogen, has been known to impair non-specific and specific immunity. Macrophages play an important role in host defense against tumors and microorganisms and a number of compounds are implicated in macrophage cytotoxicity. Since activated by the reaction of LPS with CD14, macrophages produce nitric oxide (NO) that is a cytotoxic effector molecule in cell killing. In the present study, we investigated whether the alteration of CD14 level on macrophages by AFB(1) affects NO production in murine peritoneal macrophages. When macrophages were stimulated with LPS after AFB(1)-pretreatment, or they were co-treated with LPS and AFB(1), the NO production decreased in a dose-dependent manner. In contrast, when macrophages were post-treated with AFB(1) after LPS-stimulation, NO production was unchanged. DNA, RNA, and protein synthesis were reduced by AFB(1)-pretreatment of macrophages. The addition of anti-CD14 antibodies to the cultures decreased NO production further. FACS analysis showed that the binding of anti-CD14 antibodies to the macrophages was suppressed by AFB(1)-pretreatment followed by LPS-stimulation. However, AFB(1) does not alter the binding anti-CD14 antibodies to the macrophages without LPS-stimulation. In contrast, AFB(1) pretreatment increased an amount of CD14 released in culture medium. Taken together, these data indicate that the reduced NO production in murine peritoneal macrophages by AFB(1)-pretreatment is related to the suppressed expression of CD14 on macrophage membrane and to the increased secretion of it to culture medium after LPS-stimulation.
Moon
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College of Pharmacy, Sungkyunkwan University, 300, Chunchun-dong Suwon City, South Korea.
Pyo
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D016604:Aflatoxin B1 / Q000633:toxicity*; D000818:Animals; D018950:Lipopolysaccharide Receptors / Q000502:physiology*; D008070:Lipopolysaccharides / Q000378:metabolism / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis*
D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D009569:Nitric Oxide; D016604:Aflatoxin B1