Figure 11.

Macrophage polarization, lipid mediator pathway induction, and LM production. (A) Polarization of macrophages in vitro. Macrophage cells (J774A.1) were polarized by treating with M1 inducer (IFN-γ at 20 ng/ml plus LPS at 100 ng/ml) or M2 inducer (IL-4 at 20 ng/ml) for the indicated time. Cells were lysed and analyzed by Immunoblotting against M1 markers CD68 and CD86, M2 markers CD163 and CD206, or β-actin (loading control). Representative image was shown from 3 different experiments. The relative amount of each protein was expressed as % of vehicle control and quantification was shown as mean ± SEM. (B) Expression of ALOX pathway enzymes in M1 or M2 macrophages. M1 and M2 cells were analyzed by immunoblotting against ALOX5, ALOX5AP, ALOX15, or β-actin (loading control). Representative image was shown from 3 different experiments and quantification was shown as mean ± SEM (n = 3). (C) Effect of MWCNTs on LM production from M1 or M2 macrophages. Macrophages were polarized with M1 or M2 inducer for 3 days. Polarized cells were then treated with DM or MWCNTs (2.5 μg/ml) for 1, 2, or 3 days. The cell-free culture media were collected for quantification of LTB4 (a), PGE2 (b), or RvD1 (c) using ELISA (n = 6). *p < 0.05; **p < 0.01; ***p < 0.001.