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Volume 272, Number 2, Issue of January 10, 1997 pp. 1402-1411
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Nitric Oxide Protects Cultured Rat Hepatocytes from Tumor Necrosis Factor-alpha -induced Apoptosis by Inducing Heat Shock Protein 70 Expression

(Received for publication, April 22, 1996, and in revised form, August 28, 1996)

Young-Myeong Kim Dagger § , Michael E. de Vera Dagger , Simon C. Watkins par and Timothy R. Billiar Dagger

From the Departments of Dagger  Surgery, § Pharmacology, and par  Cell Biology and Physiology, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15261

Nitric oxide (NO) and tumor necrosis factor-alpha (TNFalpha ) play important roles in the pathogenesis of liver disease during acute inflammation. The present study was designed to elucidate the effect of NO pre-exposure on TNFalpha -induced hepatotoxicity. Pretreatment of primary cultures of rat hepatocytes with the NO donor S-nitroso-N-acetylpenicillamine (SNAP) induced the expression of heat shock protein 70 (HSP70) mRNA and protein, which was associated with thermotolerance and cytoprotection from TNFalpha +actinomycin D-induced hepatotoxicity and apoptosis. SNAP transiently changed the intracellular redox state by inducing glutathione (GSH) oxidation associated with the formation of S-nitrosoglutathione (GSNO). HSP70 mRNA was also induced by the GSH-oxidizing agent diamide and the GSH-conjugating agent N-ethylmaleimide, suggesting that NO induces HSP70 expression through GSH oxidation.

The protective effect of SNAP pretreatment on TNFalpha -induced apoptosis correlated with the level of HSP70 expression. SNAP pretreatment inhibited reactive oxygen intermediate generation and lipid peroxidation effects that were reversed by blocking HSP70 expression using an antisense oligonucleotide to HSP70. Finally, endogenous NO formation, induced in hepatocytes stimulated with interferon-gamma and interleukin-1beta , led to the formation of GSNO and GSSG, induced HSP70, and attenuated TNFalpha -mediated cytotoxicity. These findings demonstrated that NO can induce resistance to TNFalpha -induced hepatotoxicity, possibly through the stimulation of HSP70 expression.


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Y.-M. Kim, H.-T. Chung, R. L. Simmons, and T. R. Billiar
Cellular Non-heme Iron Content Is a Determinant of Nitric Oxide-mediated Apoptosis, Necrosis, and Caspase Inhibition
J. Biol. Chem., April 6, 2000; 275(15): 10954 - 10961.
[Abstract] [Full Text] [PDF]


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Mol. Pharmacol.Home page
H. G. Seo, T. Nishinaka, and C. Yabe-Nishimura
Nitric Oxide Up-Regulates Aldose Reductase Expression in Rat Vascular Smooth Muscle Cells: A Potential Role for Aldose Reductase in Vascular Remodeling
Mol. Pharmacol., April 1, 2000; 57(4): 709 - 717.
[Abstract] [Full Text]


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GutHome page
C L Potter and P J Hanson
Exogenous nitric oxide inhibits apoptosis in guinea pig gastric mucous cells
Gut, February 1, 2000; 46(2): 156 - 162.
[Abstract] [Full Text] [PDF]


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J. Cell Sci.Home page
Z Dastoor and J Dreyer
Nuclear translocation and aggregate formation of heat shock cognate protein 70 (Hsc70) in oxidative stress and apoptosis
J. Cell Sci., January 8, 2000; 113(16): 2845 - 2854.
[Abstract] [PDF]


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Infect. Immun.Home page
K. B. Seydel, S. J. Smith, and S. L. Stanley Jr.
Innate Immunity to Amebic Liver Abscess Is Dependent on Gamma Interferon and Nitric Oxide in a Murine Model of Disease
Infect. Immun., January 1, 2000; 68(1): 400 - 402.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Gastrointest. Liver Physiol.Home page
J. Plumpe, N. P. Malek, C.-T. Bock, T. Rakemann, M. P. Manns, and C. Trautwein
NF-kappa B determines between apoptosis and proliferation in hepatocytes during liver regeneration
Am J Physiol Gastrointest Liver Physiol, January 1, 2000; 278(1): G173 - G183.
[Abstract] [Full Text] [PDF]


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Hum Exp ToxicolHome page
A Akbay, K Cinar, O Uzunalimoglu, S Eranil, C Yurdaydin, H Bozkaya, and M Bozdayi
Serum cytotoxin and oxidant stress markers in N-acetylcysteine treated thioacetamide hepatotoxicity of rats
Human and Experimental Toxicology, November 1, 1999; 18(11): 669 - 676.
[Abstract] [PDF]


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FASEB J.Home page
C. DIDIER, N. EMONET-PICCARDI, J.-C. BéANI, J. CADET, and M.-J. RICHARD
L-arginine increases UVA cytotoxicity in irradiated human keratinocyte cell line: potential role of nitric oxide
FASEB J, October 1, 1999; 13(13): 1817 - 1824.
[Abstract] [Full Text]


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Mol. Pharmacol.Home page
K. B. Sandau, D. Callsen, and B. Brüne
Protection against Nitric Oxide-Induced Apoptosis in Rat Mesangial Cells Demands Mitogen-Activated Protein Kinases and Reduced Glutathione
Mol. Pharmacol., October 1, 1999; 56(4): 744 - 751.
[Abstract] [Full Text]


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J. Neurosci.Home page
Y.-M. Kim, H.-T. Chung, S.-S. Kim, J.-A Han, Y.-M. Yoo, K.-M. Kim, G.-H. Lee, H.-Y. Yun, A. Green, J. Li, et al.
Nitric Oxide Protects PC12 Cells from Serum Deprivation-Induced Apoptosis by cGMP-Dependent Inhibition of Caspase Signaling
J. Neurosci., August 15, 1999; 19(16): 6740 - 6747.
[Abstract] [Full Text] [PDF]


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Cardiovasc ResHome page
R. D Rakhit, R. J Edwards, and M. S Marber
Nitric oxide, nitrates and ischaemic preconditioning
Cardiovasc Res, August 15, 1999; 43(3): 621 - 627.
[Full Text] [PDF]


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Infect. Immun.Home page
K. Guzik, J. Dobrucki, and J. Pryjma
Heat-Shocked Monocytes Are Resistant to Staphylococcus aureus-Induced Apoptotic DNA Fragmentation due to Expression of HSP72
Infect. Immun., August 1, 1999; 67(8): 4216 - 4222.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
K. Furuke, P. R. Burd, J. A. Horvath-Arcidiacono, K. Hori, H. Mostowski, and E. T. Bloom
Human NK Cells Express Endothelial Nitric Oxide Synthase, and Nitric Oxide Protects Them from Activation-Induced Cell Death by Regulating Expression of TNF-{alpha}
J. Immunol., August 1, 1999; 163(3): 1473 - 1480.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
C. Sciorati, P. Rovere, M. Ferrarini, C. Paolucci, S. Heltai, R. Vaiani, E. Clementi, and A. A. Manfredi
Generation of Nitric Oxide by the Inducible Nitric Oxide Synthase Protects {gamma}{delta} T Cells from Mycobacterium tuberculosis-Induced Apoptosis
J. Immunol., August 1, 1999; 163(3): 1570 - 1576.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
K. Wright, G. Kolios, J. Westwick, and S. G. Ward
Cytokine-induced Apoptosis in Epithelial HT-29 Cells Is Independent of Nitric Oxide Formation. EVIDENCE FOR AN INTERLEUKIN-13-DRIVEN PHOSPHATIDYLINOSITOL 3-KINASE-DEPENDENT SURVIVAL MECHANISM
J. Biol. Chem., June 11, 1999; 274(24): 17193 - 17201.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
J. Li, C. A. Bombeck, S. Yang, Y.-M. Kim, and T. R. Billiar
Nitric Oxide Suppresses Apoptosis via Interrupting Caspase Activation and Mitochondrial Dysfunction in Cultured Hepatocytes
J. Biol. Chem., June 11, 1999; 274(24): 17325 - 17333.
[Abstract] [Full Text] [PDF]


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International Journal of ToxicologyHome page
I. Latour and P. Buc-Calderon
Survival and Metabolic Function of Freshly Isolated Rat Hepatocytes Exposed First to a Heat Shock and Then to an Oxidative Stress
International Journal of Toxicology, June 1, 1999; 18(4): 239 - 244.
[Abstract] [PDF]


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Am. J. Physiol. Gastrointest. Liver Physiol.Home page
J. Li and T. R. Billiar
IV. Determinants of nitric oxide protection and toxicity in liver
Am J Physiol Gastrointest Liver Physiol, May 1, 1999; 276(5): G1069 - G1073.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
H. Li, Z. M. Marshall, and A. R. Whorton
Stimulation of cystine uptake by nitric oxide: regulation of endothelial cell glutathione levels
Am J Physiol Cell Physiol, April 1, 1999; 276(4): C803 - C811.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
L. Rossig, B. Fichtlscherer, K. Breitschopf, J. Haendeler, A. M. Zeiher, A. Mulsch, and S. Dimmeler
Nitric Oxide Inhibits Caspase-3 by S-Nitrosation in Vivo
J. Biol. Chem., March 12, 1999; 274(11): 6823 - 6826.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
A. Morikawa, Y. Kato, T. Sugiyama, N. Koide, D. Chakravortty, T. Yoshida, and T. Yokochi
Role of Nitric Oxide in Lipopolysaccharide-Induced Hepatic Injury in D-Galactosamine-Sensitized Mice as an Experimental Endotoxic Shock Model
Infect. Immun., March 1, 1999; 67(3): 1018 - 1024.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Gastrointest. Liver Physiol.Home page
C. H. Lang, G. J. Nystrom, and R. A. Frost
Regulation of IGF binding protein-1 in Hep G2 cells by cytokines and reactive oxygen species
Am J Physiol Gastrointest Liver Physiol, March 1, 1999; 276(3): G719 - G727.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
Y.-M. Kim, C. A. Bombeck, and T. R. Billiar
Nitric Oxide as a Bifunctional Regulator of Apoptosis
Circ. Res., February 19, 1999; 84(3): 253 - 256.
[Full Text] [PDF]


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CirculationHome page
J. Koglin, D. J. Granville, T. Glysing-Jensen, J. S. Mudgett, C. M. Carthy, B. M. McManus, and M. E. Russell
Attenuated Acute Cardiac Rejection in NOS2 -/- Recipients Correlates With Reduced Apoptosis
Circulation, February 16, 1999; 99(6): 836 - 842.
[Abstract] [Full Text] [PDF]


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Circ. Res.Home page
D. J. Ing, J. Zang, V. J. Dzau, K. A. Webster, and N. H. Bishopric
Modulation of Cytokine-Induced Cardiac Myocyte Apoptosis by Nitric Oxide, Bak, and Bcl-x
Circ. Res., January 22, 1999; 84(1): 21 - 33.
[Abstract] [Full Text] [PDF]


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EndocrinologyHome page
A. L. Scarim, M. R. Heitmeier, and J. A. Corbett
Heat Shock Inhibits Cytokine-Induced Nitric Oxide Synthase Expression by Rat and Human Islets
Endocrinology, December 1, 1998; 139(12): 5050 - 5057.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
Y.-M. Kim, T.-H. Kim, D.-W. Seol, R. V. Talanian, and T. R. Billiar
Nitric Oxide Suppression of Apoptosis Occurs in Association with an Inhibition of Bcl-2 Cleavage and Cytochrome c Release
J. Biol. Chem., November 20, 1998; 273(47): 31437 - 31441.
[Abstract] [Full Text] [PDF]


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Infect. Immun.Home page
S. Liu, L. S. Khemlani, R. A. Shapiro, M. L. Johnson, K. Liu, D. A. Geller, S. C. Watkins, S. M. Goyert, and T. R. Billiar
Expression of CD14 by Hepatocytes: Upregulation by Cytokines during Endotoxemia
Infect. Immun., November 1, 1998; 66(11): 5089 - 5098.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Gastrointest. Liver Physiol.Home page
B. E. Jones and M. J. Czaja
III. Intracellular signaling in response to toxic liver injury
Am J Physiol Gastrointest Liver Physiol, November 1, 1998; 275(5): G874 - G878.
[Abstract] [Full Text] [PDF]


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J. Immunol.Home page
Y.-M. Kim, R. V. Talanian, J. Li, and T. R. Billiar
Nitric Oxide Prevents IL-1{beta} and IFN-{gamma}-Inducing Factor (IL-18) Release from Macrophages by Inhibiting Caspase-1 (IL-1{beta}-Converting Enzyme)
J. Immunol., October 15, 1998; 161(8): 4122 - 4128.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Lung Cell. Mol. Physiol.Home page
G. D. Ceneviva, E. Tzeng, D. G. Hoyt, E. Yee, A. Gallagher, J. F. Engelhardt, Y.-M. Kim, T. R. Billiar, S. A. Watkins, and B. R. Pitt
Nitric oxide inhibits lipopolysaccharide-induced apoptosis in pulmonary artery endothelial cells
Am J Physiol Lung Cell Mol Physiol, October 1, 1998; 275(4): L717 - L728.
[Abstract] [Full Text] [PDF]


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Am. J. Physiol. Cell Physiol.Home page
Y. Xu, S. Bialik, B. E. Jones, Y. Iimuro, R. N. Kitsis, A. Srinivasan, D. A. Brenner, and M. J. Czaja
NF-kappa B inactivation converts a hepatocyte cell line TNF-alpha response from proliferation to apoptosis
Am J Physiol Cell Physiol, October 1, 1998; 275(4): C1058 - C1066.
[Abstract] [Full Text] [PDF]


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Proc. Natl. Acad. Sci. USAHome page
S. Ambs, M. O. Ogunfusika, W. G. Merriam, W. P. Bennett, T. R. Billiar, and C. C. Harris
Up-regulation of inducible nitric oxide synthase expression in cancer-prone p53 knockout mice
PNAS, July 21, 1998; 95(15): 8823 - 8828.
[Abstract] [Full Text] [PDF]


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J. Biol. Chem.Home page
D. Sekkai, F. Aillet, N. Israel, and M. Lepoivre
Inhibition of NF-kappa B and HIV-1 Long Terminal Repeat Transcriptional Activation by Inducible Nitric Oxide Synthase 2 Activity
J. Biol. Chem., February 13, 1998; 273(7): 3895 - 3900.
[Abstract] [Full Text] [PDF]


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International Journal of ToxicologyHome page
R. S. Mangipudy, P. S. Rao, A. Andrews, T. J. Bucci, F. A. Witzmann, and H. M. Mehendale
Dose-Dependent Modulation of Cell Death: Apoptosis Versus Necrosis in Thioacetamide Hepatotoxicity
International Journal of Toxicology, February 1, 1998; 17(2): 193 - 211.
[Abstract] [PDF]


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FASEB J.Home page
P. Rauhala and C. C. Chiueh
Neuroprotection by S-nitrosoglutathione of brain dopamine neurons from oxidative stress
FASEB J, February 1, 1998; 12(2): 165 - 173.
[Abstract] [Full Text]


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J CARDIOVASC PHARMACOL THERHome page
R.J. Moen, K. P. LaVoi, M. Zhang, and M. J. Blake
Clonidine-Induced Heat-Shock Protein Expression in Rat Aorta
Journal of Cardiovascular Pharmacology and Therapeutics, January 1, 1998; 3(2): 171 - 184.
[Abstract] [PDF]


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Circ. Res.Home page
S. Dimmeler, V. Rippmann, U. Weiland, J. Haendeler, and A. M. Zeiher
Angiotensin II Induces Apoptosis of Human Endothelial Cells : Protective Effect of Nitric Oxide
Circ. Res., December 19, 1997; 81(6): 970 - 976.
[Abstract] [Full Text]


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J. Biol. Chem.Home page
Y.-M. Kim, R. V. Talanian, and T. R. Billiar
Nitric Oxide Inhibits Apoptosis by Preventing Increases in Caspase-3-like Activity via Two Distinct Mechanisms
J. Biol. Chem., December 5, 1997; 272(49): 31138 - 31148.
[Abstract] [Full Text] [PDF]




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