Ethanol modulates induction of nitric oxide synthase in glial cells by endotoxin

Although ethanol has long been recognized as an immunosuppressant, the effects of ethanol on immune functions in the central nervous system (CNS) have not been well characterized. Glial cells function as immune effector cells within the CNS. Nitric oxide (NO), generated by inducible NO synthase (iNOS) of activated glial cells, appears to participate in the immune defense and the pathogenesis of brain injury and several neurologic diseases. The goal of the present study was to examine the effects of ethanol on NO production and mRNA expression of iNOS following its induction by bacterial endotoxin lipopolysaccharide (LPS) in cultured glial cells. After incubation of mixed glia with LPS for 24 hr, the levels of nitrite in the culture medium were assayed by Griess reaction. We found that LPS (10-500 ng/ml) induced a concentration-dependent increase in the production of NO which was abolished by the selective iNOS inhibitor aminoguanidine. While ethanol treatment (25 to 400 mM, 24 hr exposure) had no direct effect on basal NO production, it significantly suppressed the LPS-induced increase of nitrite levels in a concentration-dependent manner. Using a semiquantitative reverse transcriptase polymerase chain reaction, we found that while ethanol by itself was unable to induce iNOS mRNA, it nevertheless suppressed LPS-induced iNOS mRNA expression. Our results that ethanol had no direct effect on NO production but inhibited LPS-induced NO, indicated an immunomodulatory role by ethanol. These findings suggest that ethanol may ameliorate the consequences of overwhelming NO generation through iNOS induction in glial cells following infection, inflammation or CNS injuries.