Figure 11.

Pirfenidone regulates fibrocyte migration. Fibrocytes were cultured and CD45-positive cells were selected using magnetic beads. (A) The percentage of CD45 and collagen-I (Col-I) double-positive cells in the lung culture after magnetic selection was 75.8% on fluorescence-activated cell sorter analysis. A representative figure from 4 separate experiments is shown. (B) Fibrocyte (CD45-positive mesenchymal cell) chemotaxis was examined using a Boyden chamber assay system. Medium alone or medium containing chemokine (CC motif) ligand-2 (CCL2; 100, 200, and 500 ng/mL) was added to the lower chamber, and fibrocytes with or without pirfenidone (100 μg/mL) were added to the upper chamber. After incubation overnight at 37°C in a moist 5% CO₂/95% air atmosphere incubator, transmigration cells were counted. CCL2 at 200 and 500 ng/mL stimulated fibrocyte migration, which was significantly attenuated by pirfenidone (P = 0.0232 and 0.0025, respectively). Data are means ± standard error from 4 separate experiments, each assayed for chemotaxis in triplicate. PFD, pirfenidone.