The aim of the study: to examine the anti-inflammatory effect of lithium chloride by exposing the human neutrophils to serum of patients with septic shock in vitro.Material and methods. The study was carried out on neutrophils extracted from the blood of 6 healthy donors, which were activated with serum from patients with septic shock. The neutrophil activity was evaluated with fluorescent antibodies to the CD11b and CD66b markers of degranulation. The level of human neutrophil apoptosis and necrosis was assessed 22 hours after extraction; quantitative assessment was made using annexin V and propidium iodide with flow cytofluorimetry. Intact and activated neutrophils were treated with 0.3, 3.0 and 9.0 mmol lithium chloride solution.Results. The level of CD11b expression on the surface of intact neutrophils (healthy donors) was 3434.50 [3311.0-3799.0] arbitrary fluorescence units (AFU). Incubation of neutrophils with serum of patients with septic shock increased CD11b expression 2.5 times to 8589.0 [7279.0-11258.0] AFU (P=0.005) vs intact leukocytes, and increased CD66b expression 2.7 times up to 27 600.0 [22 999.0-28 989.0] AFU ((P=0.005) vs intact neutrophils. Lithium chloride in concentrations of 0.3, 3.0 and 9.0 mmol in a dose-dependent manner reduced the level of expression of CD11b and CD66b molecules on the surface of activated neutrophils. Septic serum reduced spontaneous neutrophil apoptosis, and 3.0 mmol and higher lithium chloride solution induced spontaneous neutrophil apoptosis.Conclusion. Lithium chloride reduces the activation of neutrophils preactivated by serum of patients with septic shock, reduces expression of CD11b and CD66b molecules on the neutrophil surface, inhibiting the process of their activation (degranulation). Lithium chloride in concentration of 3.0 mmol and higher is able to induce spontaneous apoptosis of neutrophils activated by serum of patients with septic shock.
Objectives. The goal is to study the effect of lithium chloride on the intensity of endotheliocytes apoptosis in a monolayer in vitro under the action of blood serum of patients with the syndrome of systemic inflammatory response in severe multiple trauma. Materials and methods. We used toxic blood serum of 5 patients with severe multiple trauma. As controls we used blood serum of 5 healthy donors. In different series of the experiment EA.hy926 endothelial cells were incubated with blood serum of a healthy person (control), with blood serum of patients with systemic inflammatory response syndrome in severe multiple trauma. Lithium chloride was added to the cell samples at final concentrations of 0.01, 0.1, 1, 10 mmol/L. After incubation the cells were removed with trypsin-versen solution fixed with 70 % ethanol and stained with propidium iodide. Cells containing fragmented genomic DNA were analyzed by flow cytometry. Results. It was revealed that toxic serum suppressed GSK-3p phosphorylation in endotheliocytes and also caused the splitting of VE-cadherin, a decrease in the amount of claudine and actin, initiating the destruction of intercellular contacts of the endothelial monolayer and apoptosis of endotheliocytes. Incubation of a monolayer of endotheliocytes with lithium chloride at a concentration of 1.0 mmol/l and higher almost completely prevented the dismantling of claudine, actin and VE-cadherin, and also reduced the intensity of apoptosis of endotheliocytes by more than 2 times. It was found that preincubation with lithium chloride at a concentration of 1 mmol/L not only prevented the inactivation of GSK-Зр, but even stimulated its phosphorylation. Conclusion. Lithium chloride prevents the dismantling of claudine and VE-cadherin in the intercellular contacts, reduces the number of apoptotic cells in the monolayer of the endothelial cells of the EA.hy926 line under the action of the blood serum of patients with a systemic inflammatory response syndrome in severe multiple trauma, which may indicate a protective effect of the drug on endothelial barrier. The results of this investigation suggest that the protective effect of lithium chloride on endothelium is realized via GSK-3p phosphorylation.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.