Hypertonicity induces 5-LO product formation and p38 MAPK activation.
(A) To determine 5-LO product formation, NaCl was added to freshly isolated PMNLs (5 × 106) in PGC buffer to a final volume of 1 mL, 3 minutes prior to stimulation with 40 μM AA. After 5 minutes at 37°C, 5-LO product formation was determined. Results are given as mean + SE (n = 3). Studentt test; **P < .01. (B) To determine the activation of p38 MAPK, NaCl was added to freshly isolated PMNLs (5 × 106) in PGC buffer to a final volume of 100 μL. After 3 minutes at 37°C, incubations were terminated and samples were analyzed for dually phosphorylated p38 MAPK by immunoblotting (upper panel); equal amounts of protein were evaluated with anti-p38 MAPK antibodies (lower panel). Results are representative of at least 3 separate experiments.
