Prolonged exposure to intermittent alcohol vapors decreases the ACTH as well as hypothalamic nitric oxide and cytokine responses to endotoxemia

Abstract

Background: Prolonged exposure to alcohol blunts the response of the hypothalamic-pituitary-adrenal (HPA) axis to various stressors, including the systemic injection of a lipopolysaccharide (LPS). We previously showed that decreased synthesis of the hypothalamic peptides corticotropin-releasing factor (CRF) and vasopressin (VP) played a central role in this phenomenon. However, the mechanisms that lead to decreased hypothalamic neuronal activity have not been identified. In the present work, we tested the hypothesis that alcohol decreased signals that are elicited by LPS and that stimulate hypothalamic CRF and VP synthesis, namely nitric oxide (NO) and the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6).

Methods: Adult male rats were exposed to intermittent (5 hr/day) alcohol vapors for 5 days. Control animals were kept in comparable chambers but not exposed to the vapors. On day 6, the animals received an injection of LPS through permanent indwelling intravenous cannulae. The dependent variables were plasma ACTH levels measured by IRMA (immunoradiometric assay); pituitary and hypothalamic TNF-alpha and IL-6 mRNA levels measured by RNase protection assay; basal activity of neuronal NO synthase measured by conversion of [14C]arginine to [14C]citrulline, the constitutive enzyme that synthesizes NO and modulates the influence of this gas on LPS-induced HPA axis activity; and basal and LPS-induced levels of citrulline (an index of NO formation) in the hypothalamus, measured by immunocytochemistry.

Results: After injection with LPS, rats that were pretreated with alcohol exhibited a significantly (p < 0.01) decreased release of ACTH, compared with controls. There was no difference in basal NO synthase activity or hypothalamic citrulline levels. In contrast, LPS-induced hypothalamic citrulline levels were significantly (p < 0.01) lower in alcohol-exposed rats, as were pituitary TNF-alpha and IL-6 transcripts. In the hypothalamus, the TNF-alpha but not IL-6 response to LPS was also reduced.

Conclusions: These results indicate that prolonged exposure to alcohol decreases the ACTH, hypothalamic NO and TNF-alpha, and pituitary TNF-alpha and IL-6 responses to LPS. This suggests that altered NO and proinflammatory cytokine levels in the brain may modulate the inhibitory influence exerted by alcohol on the HPA axis response to endotoxemia.