Cell-Specific Effects of Ethanol on Nitric Oxide Synthase Induced by Inflammatory Mediators

DOI: 10.4236/pp.2014.513132   PDF   HTML   XML   2,567 Downloads   3,118 Views  


To examine the pro- and anti-oxidant properties of nitric oxide (NO) production in alcoholic liver disease, we compared the effects of ethanol pretreatment (24 hrs, 100 mM concentration in a dedicated CO2 incubator) on the induction of inducible NO synthase and its activity in a cell culture system. We employed two types of liver cells as models for the intact liver parenchyma: the rat hepatoma cell FTO2B and rat hepatocytes in primary culture. Cells were incubated with a combination of cytokines (IFN-γ, TNF-α, IL-β) and LPS in the presence or absence of (85 - 93 mM) ethanol in the culture media. At series of time intervals, production of nitric oxide was measured as the accumulation of nitrite and nitrate, using Griess assay. The results revealed that 1) total NO formation was attenuated by ethanol in hepatocytes (ca. 50%) but augmented in FTO2B cells (ca. 37%) and 2) both pretreatment and co-treatment with ethanol were necessary for maximal ethanol effect. These results indicate that the effects of ethanol on inflammatory processes, such as induction of NO synthesis, are cell-type specific.

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O. Obih, P. and Potter, B. (2014) Cell-Specific Effects of Ethanol on Nitric Oxide Synthase Induced by Inflammatory Mediators. Pharmacology & Pharmacy, 5, 1202-1208. doi: 10.4236/pp.2014.513132.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] Center for Disease Control (CDC) (2014) Alcohol and Public Health: Fact Sheets—Alcohol.
[2] Cederbaum, A.I. (2012) CYP2E1 Potentiates Toxicity in Obesity and after Chronic Ethanol Treatment. Drug Metabolism and Drug Interaction, 27, 125-144.
[3] Baraona, E., Zeballos, G.A., Shoichet, L., Mak, K.M. and Lieber, S.S. (2002) Ethanol Consumption Increases Nitric Oxide Production in Rats and Its Peroxynitrite-Mediated Toxicity Is Attenuated by Polyenylphosphatidylcholine. Alcoholism: Clinical and Experimental Research, 26, 883-889.
[4] Nanji, A.A. and Hiller-Sturmhöfel, S. (1997) Apoptosis and Necrosis: Two Types of Cell Death in Alcoholic Liver Disease. Alcohol Health and Research World, 21, 325-330.
[5] Adams, A.G. and Jourdan, C. (1984) Infection in Alcoholic. Medical Clinics of North America, 68, 179-200.
[6] Choi, J. (2012) Oxidative Stress, Endogenous Antioxidants, Alcohol, and Hepatitis C: Pathogenic Interactions and Therapeutic Considerations. Free Radical Biology and Medicine, 52, 1135-1150.
[7] Cederbaum, A.I. (2001) Introduction—Serial Review: Alcohol, Oxidative Stress and Cell Injury. Free Radical Biology and Medicine, 31, 1524-1526.
[8] Joshi, M.S., Ponthier, J.L. and Lancaster Jr., J.R. (1999) Cellular Antioxidant and Pro-Oxidant Actions of Nitric Oxide. Free Radical Biology and Medicine, 27, 1357-1366.
[9] Cederbaum, A.I. (2014) Cyrochrome P450 2E1, Oxidative Stress, JNK, and Autophagy in Acute Alcohol-Induced Fatty Liver. In: Hayat, M. A., Ed., Autophagy: Cancer, Other Pathologies, Inflammation, Immunity, Infection, and Aging, Volume 2: Role in General Diseases, Academic Press, Waltham, 341-356.
[10] Wu, D. and Cederbaum, A.I. (2003) Alcohol, Oxidative Stress, and Free Radical Damage. Alcohol Research and Health, 27, 277-284.
[11] Zhang, H. and Potter, B.J. (1992) The Effect of Ethanol Metabolism on Ferritin Uptake by Freshly Isolated Rat Hepatocytes: Is Acetaldehyde Responsible for This Alteration? Alcoholism: Clinical and Experimental Research, 16, 301-307.
[12] Potter, B.J., Blades, B., McHUgh, T.A., Nunes, R.M., Beloqui, O., Slott, P.A. and Rand, J.H. (1989) Effects of Endotoxin on Iron Uptake by the Hepatocyte. American Journal of Physiology, 257, G524-G531.
[13] Kim, Y.M., Bergonia, H. and Lancaster Jr., J.R. (1995) Nitrogen Oxide-Induced Autoprotection in Isolated Rat Hepatocytes. FEBS Letters, 374, 228-232.
[14] Jin, M., Ande, A., Kumar, A. and Kumar, S. (2013) Regulation of Cytochrome P450 2E1 Expression by Ethanol: Role of Oxidative Stress-Mediated PKC/JNK/SP1 Pathway. Cell Death and Disease, 4, e554.
[15] Yang, S.U., Lee, J.H. and Park, J. (2008) Ethanol Induces Peroxynitrite-Mediated Toxicity through Inactivation of NADP+-Dependent Isocitrate Dehydrogenase and Superoxide Dismutase. Biochimie, 90, 1316-1324.
[16] Griffon, B., Chevanne, M., Morel, P., Cillard, P. and Sergent, O. (2000) Activated Macrophages Increase the Susceptibility of Rat Hepatocytes to Ethanol-Induced Oxidative Stress: Conflicting Effects of Nitric Oxide. Alcohol and Alcoholism, 35, 230-235.
[17] Wang, E., Spitzer, J.J. and Chamulirat, W. (1999) Differential Regulation of Inducible Nitric Oxide Synthase Gene Expression by Ethanol in the Human Intestinal Epithelial Cell Line DLD-1. Nitric Oxide: Biology and Chemistry, 3, 244-253.
[18] Spitzer, J.A. and Spitzer, J.J. (2000) Lipopolysaccharide Tolerance and Ethanol Modulate Hepatic Nitric Oxide Production in Gender-Dependent Manner. Alcohol, 21, 27-35.
[19] Zuckerman, S.H., Bryan-Poole, N., Evans, G.F., Short, L. and Glasebrook, A.L. (1995) In Vivo Modulation of Murine Serum Tumour Necrosis Factor and Interleukin-6 Levels during Endotoxemia by Oestrogen Agonists and Antagonists. Immunology, 86, 18-24.
[20] Moezi, L., Heidari, R., Amirghofran, Z., Nekooeian, A.A., Monabati, A. and Dehpour, A.R. (2013) Enhanced Anti-Ulcer Effect of Pioglitazone on Gastric Ulcers in Cirrhotic Rats: The Role of Nitric Oxide and IL-1β. Pharmacological Reports, 651, 134-143.

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