Role of calcium in tumor necrosis factor-alpha production by activated macrophages.
The role of calcium in the production of tumor necrosis factor-alpha (TNF-alpha) by the lipopolysaccharide (LPS)-stimulated macrophage cell line, J774.1, was investigated. Flow cytometric measurement of intracellular calcium concentration ([Ca]i) using 2-(3,6-bis(acetyl-oxy)-2,7-dichloro-9H-xanthen-9-yl)benzoic acid (fluo-3)-loaded J774.1 cells revealed that LPS at more than 0.1 microgram/ml increased [Ca]i transiently in the presence or absence of serum, and that a mixture of a calcium chelator, ethyleneglycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), and a calcium release blocker from intracellular store sites, 8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate hydrochloride (TMB-8), inhibited the [Ca]i response induced by LPS. In concordance with this, production of TNF-alpha was inhibited by EGTA and/or TMB-8. These reagents also reduced the level of TNF-alpha mRNA significantly. These results indicate that the transient increase of [Ca]i plays a role in LPS-induced expression of TNF-alpha by the macrophage cell line.
Watanabe
N
N
Department of Biomolecular Science, Faculty of Science, Toho University, Chiba.
Suzuki
J
J
Kobayashi
Y
Y
D000818:Animals; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D004533:Egtazic Acid / Q000494:pharmacology; D005707:Gallic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008262:Macrophage Activation; D008264:Macrophages / Q000378:metabolism*; D051379:Mice; D012333:RNA, Messenger / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D008070:Lipopolysaccharides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D000001:Calcimycin; D004533:Egtazic Acid; C006014:8-(N,N-diethylamino)octyl-3,4,5-trimethoxybenzoate; D005707:Gallic Acid; D002118:Calcium