Reduced Hepatotoxicity of Acetaminophen in Mice Lacking Inducible Nitric Oxide Synthase: Potential Role of Tumor Necrosis Factor-α and Interleukin-10

doi:10.1006/taap.2002.9474

Toxicology and Applied Pharmacology

Volume 184, Issue 1, 1 October 2002, Pages 27-36

https://doi.org/10.1006/taap.2002.9474 Get rights and content

Abstract

Macrophage-derived inflammatory mediators have been implicated in tissue injury induced by a number of hepatotoxicants. In the present studies, we used transgenic mice with a targeted disruption of the gene for inducible nitric oxide synthase (NOS II) to analyze the role of nitric oxide in inflammatory mediator production in the liver and in tissue injury induced by acetaminophen. Treatment of wild-type mice with acetaminophen (300 mg/kg) resulted in centrilobular hepatic necrosis, which was evident within 3 h and reached a maximum at 18 h. This was correlated with NOS II expression and nitrotyrosine staining of the liver, which was most prominent after 6 h. Expression of mRNA for tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10), matrix metalloproteinase-9, and connective tissue growth factor (CTGF) also increased in the liver following acetaminophen treatment of wild-type mice. NOS II knockout mice were found to be less sensitive to the hepatotoxic effects of acetaminophen than wild-type mice. This did not appear to be due to differences in acetaminophen-induced glutathione depletion or adduct formation. In NOS II knockout mice treated with acetaminophen, hepatic expression of TNF-α, as well as CTGF, was significantly increased compared to wild-type mice. In contrast, IL-10 expression was reduced. These data demonstrate that nitric oxide is important in hepatotoxicity induced by acetaminophen. Moreover, some of its effects may be mediated by altering production of pro- and antiinflammatory cytokines and proteins important in tissue repair.

References (48)

J.B. Bartolone et al.
Purification, antibody production, and partial amino acid sequence of the 58-kDa acetaminophen-binding liver proteins
Toxicol. Appl. Pharmacol.
(1992)
M.E. Blazka et al.
Role of proinflammatory cytokines in acetaminophen hepatotoxicity
Toxicol. Appl. Pharmacol.
(1995)
M. Bourdi et al.
Protection against acetaminophen-induced liver injury and lethality by interleukin-10: Role of inducible nitric oxide synthase
Hepatology
(2002)
M.M. Bradford
A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein dye binding
Anal. Biochem.
(1976)
A. Bruccoleri et al.
Induction of early-immediate genes by tumor necrosis factor alpha contribute to liver repair following chemical-induced hepatotoxicity
Hepatology
(1997)
S.J. Bulera et al.
Acetaminophen-arylated proteins are detected in hepatic subcellular fractions and numerous extra-hepatic tissues in CD-1 and C57Bl/6J mice
Toxicology
(1996)
T. Haruyama et al.
Regulation and significance of hepatocyte-derived matrix metalloproteinases in liver remodeling
Biochem. Biophys. Res. Commun.
(2000)
H. Ischiropoulos
Biological tyrosine nitration: A pathophysiological function of nitric oxide and reactive oxygen species
Arch. Biochem. Biophys.
(1998)
H. Ischiropoulos et al.
Peroxynitrite formation from macrophage-derived nitric oxide
Arch. Biochem. Biophys.
(1992)
T. Knittel et al.
Expression patterns of matrix metalloproteinases and their inhibitors in parenchymal and non-parenchymal cells of rat liver: Regulation by TNF-α and TGF-β1
J. Hepatol.
(1999)

P.C. Kuo et al.

Nitric oxide decreases oxidant-mediated hepatocyte injury

J. Surg. Res.

(1994)

D.L. Laskin et al.

Modulation of macrophage functioning abrogates the acute hepatotoxicity of acetaminophen

Hepatology

(1995)

D.L. Laskin et al.

Potential role of activated macrophages in acetaminophen hepatotoxicity. I. Isolation and characterization of activated macrophages from rat liver

Toxicol. Appl. Pharmacol.

(1986)

D.L. Laskin et al.

Potential role of activated macrophages in acetaminophen hepatotoxicity. II. Mechanism of macrophage accumulation and activation

Toxicol. Appl. Pharmacol.

(1986)

D.L. Laskin et al.

Hepatic nitric oxide production following acute endotoxemia in rats is mediated by increased nitric oxide synthase gene expression

Hepatology

(1995)

S.S.T. Lee et al.

Role of CYP2E1 in the hepatotoxicity of acetaminophen

J. Biol. Chem.

(1996)

S.L. Michael et al.

Acetaminophen-induced hepatotoxicity in mice lacking inducible nitric oxide synthase activity

Nitric Oxide

(2001)

L.A. Morio et al.

Distinct roles of tumor necrosis factor-α and nitric oxide in acute liver injury induced by carbon tetrachloride in mice

Toxicol. Appl. Pharmacol.

(2001)

E.E. Moussad et al.

Connective tissue growth factor: What's in a name?

Mol. Genet. Metab.

(2000)

A.K. Nussler et al.

Nitric oxide, hepatocytes and inflammation

Res. Immunol.

(1995)

K.W. Renton et al.

The prevention of acetaminophen-induced hepatotoxicity by the interferon inducer poly(rI^.rC)

Toxicol. Appl. Pharmacol.

(1984)

P.K. Smith et al.

Measurement of protein using bicinchoninic acid

Anal. Biochem.

(1985)

K. Ujike et al.

Kinetics of expression of connective tissue growth factor gene during liver regeneration after partial hepatectomy and d-galactosamine-induced liver injury in rats

Biochem. Biophys. Res. Commun.

(2000)

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Regular ArticleReduced Hepatotoxicity of Acetaminophen in Mice Lacking Inducible Nitric Oxide Synthase: Potential Role of Tumor Necrosis Factor-α and Interleukin-10

Abstract

Toxicol. Appl. Pharmacol.

Toxicol. Appl. Pharmacol.

Hepatology

Anal. Biochem.

Hepatology

Toxicology

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

J. Hepatol.

J. Surg. Res.

Hepatology

Toxicol. Appl. Pharmacol.

Toxicol. Appl. Pharmacol.

Hepatology

J. Biol. Chem.

Nitric Oxide

Toxicol. Appl. Pharmacol.

Mol. Genet. Metab.

Res. Immunol.

Toxicol. Appl. Pharmacol.

Anal. Biochem.

Biochem. Biophys. Res. Commun.

Regular Article
Reduced Hepatotoxicity of Acetaminophen in Mice Lacking Inducible Nitric Oxide Synthase: Potential Role of Tumor Necrosis Factor-α and Interleukin-10