Histamine-Induced Production of Interleukin-6 and Interleukin-8 by Human Coronary Artery Endothelial Cells Is Enhanced by Endotoxin and Tumor Necrosis Factor-α

Abstract

In this study, we tested the synergy between histamine and LPS, and histamine and TNF-α, on endothelial cell production of interleukin-6 (IL-6), interleukin-8 (IL-8), and monocyte chemoattractant protein-1 (MCP-1). Human coronary artery endothelial cells (HCAEC) were cultured in vitro with histamine (0.1 to 1000 μM) in the presence or absence of LPS or TNF-α for 24 h, and the secreted IL-6, IL-8 and MCP-1 were quantified. Unactivated HCAEC produced minimal levels of IL-6, IL-8, or MCP-1. The incubation of HCAEC with histamine resulted in low level induction of IL-6 and IL-8 production, which was dose-dependent and attained a plateau at a concentration of 10 μM. On the other hand, histamine failed to induce MCP-1 production. Stimulation of HCAEC with LPS or TNF-α caused dose-dependent increase in cytokine production. In the presence of all stimulatory concentrations of LPS and TNF-α tested, histamine was shown to further enhance IL-6 and IL-8 production. The effect of histamine on endothelial cell production of cytokines was completely inhibited by the H-1 receptor antagonist, diphenhydramine, and not by the H-2 antagonist, famotidine. Electrophoretic mobility shift assays of nuclear proteins extracted from HCAEC treated with histamine and LPS, or histamine and TNF-α, revealed amplified translocation of NF-κB proteins to the nuclei. Since both LPS and TNF-α potentiated histamine-induced cytokine production, it is possible that these activators stimulate H-1 receptor expression and/or augment the signal transduction pathways leading to the expression of IL-6 and IL-8. These results indicate the importance of synergy between histamine and other inflammatory stimuli on endothelial cell activation and implicate their cooperative participation in vascular leak and inflammation.