The blockade of ERK1/2 MAPK activation inhibited Ac-PGP production.

Abstract

<p>Human peripheral blood PMNs (5.25×10⁵) were pretreated with U0126 (10 and 20 µM) versus vehicle for 30 minutes and then incubated for 30 minutes with labeled Ac-PGP. The supernatants collected from incubated cells were placed on type I collagen (1.0 mg/ml) for 24 hours at 37°C. Ac-PGP and C¹³N¹⁵ labeled Ac-PGP were analyzing by <i>ESI-LC-MS/MS</i>. Ac-PGP values of the samples on PBS were subtracted from Ac-PGP values of samples incubated on type I collagen to determine Ac-PGP production. <b>A</b>. Detection of Ac-PGP and C¹³N¹⁵ Ac-PGP via mass spectrometry. <b>B</b>. The detection of gelatinolytic activity in culture supernatants from human PMNs stimulated with labeled Ac-PGP by g<i>elatin zymography</i> representative of three gels. <b>C</b>. The measurement of Ac-PGP production by <i>ESI-LC-MS/MS.</i> The bar graph represents the percent of relative Ac-PGP production normalized to labeled Ac-PGP control. *p<0.05 compared to labeled Ac-PGP without inhibitor pretreatment, n = 4 wells/condition.</p