Human Mast Cells (HMC-1 5C6) Enhance Interleukin-6 Production by Quiescent and Lipopolysaccharide-Stimulated Human Coronary Artery Endothelial Cells
Figure 4
Combination of H1R antagonist and protease inhibitors (PI) abrogates mast cell- (MC-) mediated potentiation of IL-6 production by LPS-stimulated endothelial cells. Endothelial cells (10,000 cells/well) were grown for 24 h in the presence of mast cells (5000 cells/well) and were activated with LPS (100 ng/mL) for a period of 24 h. To determine the effect of H1R or H2R antagonists, endothelial cells were incubated with 10 μM diphenhydramine (DPH) (a) or 10 μM famotidine (FAM) (b) for 30 min before addition of HMC-1 5C6. To determine the contribution of mast cells proteases (c), HMC-1 5C6 were pretreated with protease inhibitor (PI) cocktail overnight, washed twice, and then added to endothelial cell monolayers. After 24 h incubation, the culture supernatants were analyzed for IL-6 by ELISA. Results are mean ± SEM of quadruplicate determinations of a representative experiment. Similar results were observed in two other experiments.
