Macrophage Derived Platelet Activating Factor Implicated in the Resolution Phase of Gouty Inflammation
Figure 3
(a) Effect of recombinant human platelet activating factor acetylhydrolase (rhuPAF-AH) on TNFα release from monocytes. Freshly isolated monocytes were cultured overnight and treated with rhuPAF-AH at concentrations of 2.5, 5, 10, and 20 μg/mL, respectively, for 45 minutes prior to addition of MSU crystals for 24 hours. Afterwards, collected supernatants were analysed for TNFα by ELISA. Results are mean and SEM of 3 experiments. (b). TNFα production unmasked by costimulation of in vitro differentiated macrophages with MSU crystals and WEB2086. Monocytes were cultured in vitro for 7 days and stimulated with MSU crystals for 24 hours with or without the PAF antagonist (WEB2086). At 100 μM, coincubation with WEB2086 and urate crystals caused the release of TNFα with (Student’s paired -test). Conditioned supernatants were analysed for TNFα production by ELISA. Differentiated macrophages were also coincubated with WEB2086 at 100 μM which reduced the amount of TNFα unmasked. Values are mean and SEM of 4 experiments. D7 = supernatants from day 7 macrophages, D1 = supernatants from day 1 monocytes, and WEB = WEB2086 (PAF antagonist).
