PMID- 8700134
OWN - NLM
STAT- MEDLINE
DCOM- 19960903
LR  - 20131121
IS  - 0026-895X (Print)
IS  - 0026-895X (Linking)
VI  - 50
IP  - 2
DP  - 1996 Aug
TI  - Characterization of hepatic nitric oxide synthase: identification as the 
      cytokine-inducible form primarily regulated by oxidants.
PG  - 277-84
AB  - Induction of hepatic nitric oxide synthase (NOS) by tumor necrosis factor-alpha 
      (TNF alpha), interleukin-1 beta (IL-1 beta), interferon-gamma (IFN gamma), 
      interleukin-6 (IL-6), and lipopolysaccharide was assessed as activity and 
      immunoreactive protein. Hepatic NOS activity was cytosolic and had cofactor 
      requirements consistent with inducible nitric oxide synthase (NOS2). NOS 
      induction by TNF alpha was dose dependent from concentrations of 0.06 to 60 nM 
      and was increased 2-3-fold by IFN gamma. NOS induction was reflective of total 
      TNF alpha binding to hepatocyte receptors. Hepatocyte TNF alpha binding fit a 
      biphasic curve with high affinity (K(d) = 1.4 nM, Bmax = 3157 sites) and low 
      affinity (K(d) = 157 nM, Bmax = 204,948 sites) elements. NOS2 activity was 
      induced by lipopolysaccharide, IL-1 beta, TNF alpha, and IFN gamma but not by 
      IL-6. All cytokine stimuli were inhibited by antioxidants. Oxygen radical 
      generation was directly measured as dichlorofluoroscein fluorescence in isolated 
      mitochondria. Mitochondria from TNF alpha-treated hepatocytes generated more 
      oxygen radicals than did controls. Antioxidants reduced mitochondrial generation 
      of oxygen radicals. Activation of the transcription factor nuclear factor-kappa B 
      by TNF alpha, IFN gamma, and IL-1 beta was assessed by gel shift analysis. 
      Cytokine treatment increased nuclear factor-kappa B binding, and the addition of 
      antioxidants or rotenone inhibited cytokine activation. Taken together, these 
      data suggest that oxygen radicals, possibly generated by mitochondria, play a 
      major role in NOS2 induction by cytokines.
FAU - Duval, D L
AU  - Duval DL
AD  - Department of Environmental Health, College of Veterinary Medicine and Biomedical 
      Sciences, Colorado State University, Fort Collins 80523, USA.
FAU - Miller, D R
AU  - Miller DR
FAU - Collier, J
AU  - Collier J
FAU - Billings, R E
AU  - Billings RE
LA  - eng
GR  - DK44755/DK/NIDDK NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Mol Pharmacol
JT  - Molecular pharmacology
JID - 0035623
RN  - 0 (Antioxidants)
RN  - 0 (Cytokines)
RN  - 0 (NF-kappa B)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - Animals
MH  - Antioxidants/*pharmacology
MH  - Cells, Cultured
MH  - Cytokines/*pharmacology
MH  - Enzyme Induction/drug effects
MH  - Liver/*enzymology
MH  - Male
MH  - NF-kappa B/metabolism
MH  - Nitric Oxide Synthase/*metabolism
MH  - Oxygen/*metabolism
MH  - Rats
MH  - Rats, Sprague-Dawley
MH  - Tumor Necrosis Factor-alpha/metabolism
EDAT- 1996/08/01 00:00
MHDA- 1996/08/01 00:01
CRDT- 1996/08/01 00:00
PHST- 1996/08/01 00:00 [pubmed]
PHST- 1996/08/01 00:01 [medline]
PHST- 1996/08/01 00:00 [entrez]
PST - ppublish
SO  - Mol Pharmacol. 1996 Aug;50(2):277-84.