Effect of Sevoflurane on Activation of Human Neutrophiles in Ex Vivo Models

Abstract

The objective is to study the effect of different concentrations of sevoflurane on activation of human neutrophils in an ex vivo model.Subjects and Methods. The cell culture of venous blood neutrophils of 5 healthy men was used in this study. Neutrophil activation by lipopolysaccharide (LPS) and chemotaxis peptide N-formyl-methionine-leucine-phenylalanine (fMLP) as stimulants, was assessed by the expression level of CD11b and CD66b, IL-1β, IL-6 and IL-8, the level of phosphorylation of glycogen synthase β-kinase-3β (GSK-3β). Annexin V and propidium iodide were used to assess apoptosis. Neutrophils were exposed to 0.5, 1 and 1.5 MAC of sevoflurane to assess the effect of the drug on their activation.Results. Incubation of neutrophils with LPS and fMLP statistically significantly increased the expression of these molecules: treatment with LPS at the dose of 200 ng/ml increased CD11b and CD66b expression by 2.3 and 2.2 times (p = 0.002 and p = 0.001, respectively), while treatment with fMLP at 100 nM increased expression by 1.7 and 2.0 times (p = 0.025 and p = 0.03, respectively). When neutrophils were incubated with the same concentration of LPS after exposure to sevoflurane at a dose of 1.5 MAC, the level of CD11b and CD66b expression increased versus intact neutrophils. In this experiment, the change in CD11b expression was statistically insignificant (p = 0.055), the change in CD66b expression was statistically significant (p = 0.007). Thus, sevoflurane exposure at a dose of 1.5 MAC reduces proinflammatory activation of neutrophils induced by LPS.Conclusion. Stimulation of neutrophils by LPS was accompanied by dephosphorylation of GSK-3β, and exposure to 1.5 MAC of sevoflurane resulted in its phosphorylation. Thus, phosphorylation of GSK-3β in neutrophils by sevoflurane reduces the expression of CD11b and CD66b.