Temporal Differences in the Ability of Ethanol to Modulate Endotoxin‐Induced Increases in Inflammatory Cytokines in Muscle Under In Vivo Conditions

Abstract

Acute alcohol (EtOH) intoxication may both antagonize and potentiate the ability of monocytes/macrophages to respond to endotoxin (lipopolysaccharide [LPS]). The suppressive effects of EtOH predominate when the duration between EtOH and LPS administration is relatively short, whereas sensitization is observed under conditions when there is a relatively longer delay between EtOH and LPS exposure. Striated muscle is now recognized to possess components of both the afferent and efferent limbs of the innate immune system. The aim of the present study was to determine whether the interval between EtOH and LPS administration differentially affects the mRNA content for selected elements of the innate immune response in skeletal and cardiac muscle and to compare such changes with those occurring in liver and spleen. The content of mRNA for interleukin (IL)-6, IL-1beta, tumor necrosis factor (TNF)-alpha, and high-mobility group box (HMGB)-1, as well as toll-like receptors (TLRs)-2 and -4, were measured in gastrocnemius, heart, liver and spleen from rats orally gavaged with EtOH and then injected with LPS either two or 24 hr thereafter. EtOH intoxication two hr before LPS acutely suppressed the increased IL-6 mRNA in all tissues and antagonized the increase in plasma and tissue IL-6 protein concentration. Similarly, EtOH blunted the LPS-induced increase in tissue mRNA expression of TNF-alpha and IL-1beta. In contrast, when LPS was given 24 hr after EtOH, the increased IL-6 in striated muscle, but not in liver or spleen, was selectively potentiated. An enhanced LPS responsiveness was also observed for the late-phase cytokine HMGB1 in all tissues; however, the increased tissue expression of TNF-alpha and IL-1beta induced by LPS was not augmented. TLR4 mRNA was decreased in both heart and spleen (but unaltered in skeletal muscle and liver) of rats injected with LPS, and this change was prevented by pretreatment with EtOH. In contrast, EtOH alone increased TLR-2 mRNA content of heart, liver, and spleen but not muscle. LPS also markedly increased TLR2 mRNA in the same three tissues under control conditions, but this increase was attenuated by EtOH administered either two or 24 hr before LPS. Under in vivo conditions, the interval between EtOH exposure and LPS differentially affected the synthesis of various cytokines. In this regard, EtOH administered within two hr of LPS generally suppressed IL-6, IL-1beta, and TNF-alpha mRNAs in muscle, heart, liver, and spleen. Delaying the exposure of animals to LPS for 24 hr after EtOH, however, accentuated the increase in IL-6 and HMGB1, and for IL-6, this increased sensitivity appeared localized to striated muscle.