Figure 7.

Acute COX2 inhibition does not affect efferocytosis, but chronic or constitutive loss of COX2 activity suppresses prostanoids otherwise present across the differentiation of monocytes into BMDMs while biasing the polarization state of BMDMs toward an inflammatory phenotype. (A) BL6 PMs were culture for 7 days and then pretreated with vehicle (NT) or SC236 for 60 minutes prior to addition of labeled apMPRO cells. Numbers of apMPRO cells per PM were determined by assessing multiple micrographic fields of at least 200 PMs for 3 biological replicates. (B) Bone marrow–derived monocytes were isolated from BL6 mice and treated with vehicle (NT) or the COX2 inhibitor SC236 from time 0. Media collected on day 4 were analyzed by LC-MS/MS for eicosanoids including PGE2, the PGE2 degradation product 15keto PGE2, and the PGI2 degradation product 6ketoPGF1α. Data from 3 biological replicates. (C, D) Bone marrow–derived monocytes from BL6 and Cox2 MKO mice were treated with vehicle or SC236 from time 0 (WT = BL6 BMDM + vehicle; SC236 = BL6 BMDM + SC236; MKO = Cox2 MKO BMDM + vehicle). On day 8, BMDMs were further treated with vehicle, IL-4 (20 ng/mL), or IL-10 (20 ng/mL). On day 10, gene expression levels of the activation markers Ym1 (M2/M2a marker) and Nos2 (M1 marker) were determined by qPCR. Data are expressed as fold differences from the corresponding intragroup COX2 WT and uninhibited control (WT). ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. (A) Unpaired Student’s t test. (B–D) 2-way analysis of variance with Tukey's multiple comparisons test and adjusted P values. ns, not significant.