Suppression of Cytokine-lnduced Neutrophil Accumulation in Rat Mesenteric Venules in vivo by General Anesthesia

Most studies of neutrophil-endothelial interactions in vivo necessarily require the use of general anesthetic agents which are well known to be immunosuppressive. By using whole-mount preparations of the rat mesoappendix, we were able to study tumor necrosis factor alpha (TNF-α) induced neutrophil adhesion to the mesenteric venular endothelium in vivo without necessarily using general anesthesia. TNF-α significantly increased venular-neutrophil accumulation in a dose-dependent manner; accumulation was markedly increased at 1,2, and 4 h, but returned to baseline after 24 h. After these preliminary dose-response and time-course studies, we evaluated the influence of standard clinically effective doses of several commonly used anesthetic agents (thiopental, pentobarbital, ketamine, α-chloralose, methoxyflurane, and halothane) on the extent of neutrophil-venular accumulation induced 2 h after intraperitoneal injection of 0.4 mg/kg TNF-α, compared to unanesthetized rats. All general anesthetics tested, with the exception of methoxyflurane, significantly suppressed this response. In most cases this suppression was striking (from 60 to 85%) such that a statistically significant proinflammatory response was obscured. Although methoxyflurane also tended to suppress this response to TNF-α, it was the only agent that allowed the response to be clearly seen. Because anesthesia markedly suppresses cytokine-induced neutrophil-venular adhesion, this model should provide an important complementary technique to the classical in vivo microcirculatory approaches which do necessarily require general anesthesia.