Role of NF-kappa B in the regulation of inducible nitric oxide synthase in an MTAL cell line.
The effects of cytokines, lipopolysaccharide (LPS), 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP), and pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappa B (NF-kappa B) activation, on inducible nitric oxide synthase (iNOS) expression were studied in the medullary thick ascending limb of Henle's loop cell line ST-1. LPS + interferon-gamma (IF-gamma) promoted a time-dependent increase in nitrite (a NO metabolite) and iNOS mRNA and the appearance of NF-kappa B p50 and p65 in nuclear protein extracts. Actinomycin D but not cycloheximide prevented the LPS + IF-gamma induction of iNOS mRNA and NO synthesis, indicating that iNOS transcriptional activation by LPS + IF-gamma does not require newly synthesized proteins. PDTC inhibited the LPS + IF-gamma induction of NO, iNOS mRNA, and the appearance of NF-kappa B in nuclear protein extracts, suggesting that NF-kappa B mobilization and trans-activation of the iNOS gene mediates this induction. In contrast to other cell types, cycloheximide did not alter iNOS mRNA stability, and 8-BrcAMP did not alter basal or LPS+IF-gamma induced NO production in ST-1 cells.
Kone
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BC
Department of Medicine, University of Florida College of Medicine, Gainesville 32610-0224, USA.
Schwöbel
J
J
Turner
P
P
Mohaupt
M G
MG
Cangro
C B
CB
D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology; D004790:Enzyme Induction; D015870:Gene Expression; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008138:Loop of Henle / Q000166:cytology / Q000201:enzymology*; D051379:Mice; D008969:Molecular Sequence Data; D009088:Mucoproteins / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis* / Q000235:genetics; D015345:Oligonucleotide Probes / Q000235:genetics; D011759:Pyrrolidines / Q000494:pharmacology; D051381:Rats; D013859:Thiocarbamates / Q000494:pharmacology; D058949:Uromodulin
D016207:Cytokines; D008070:Lipopolysaccharides; D009088:Mucoproteins; D016328:NF-kappa B; D015345:Oligonucleotide Probes; D011759:Pyrrolidines; D013859:Thiocarbamates; C548612:UMOD protein, human; C548613:Umod protein, mouse; D058949:Uromodulin; D015124:8-Bromo Cyclic Adenosine Monophosphate; C020972:pyrrolidine dithiocarbamic acid; D019001:Nitric Oxide Synthase