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Pirfenidone exerts a suppressive effect on CCL18 expression in U937-derived macrophages partly by inhibiting STAT6
Yoshinobu Saito1, Arata Azuma1, Kuniko Matsuda1
1a Department of Pulmonary Medicine and Oncology , Graduate School of Medicine, Nippon Medical School , Tokyo , Japan.
Context:
CC chemokine ligand 18 (CCL18) is suggested to play a role in the development of pulmonary fibrosis. Macrophages are thought to be the main source of CCL18, and the effect of pirfenidone, an anti-fibrotic agent for idiopathic pulmonary fibrosis, on the expression of CCL18 in macrophages warrants investigation.
Objective:
The purpose of this study was to investigate the effect of pirfenidone on the expression of CCL18 in macrophages.
Materials And Methods:
U937 cells were differentiated into macrophages by phorbol myristate acetate and then stimulated with recombinant IL-4 to induce the production of CCL18. The cells were treated with pirfenidone, and the mRNA and protein levels for CCL18 were measured by a reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. The effects of pirfenidone on the IL-4 receptor (IL-4R) expression and STAT6 activation were investigated and on the JAK kinase activity were measured using the Z'-LYTE™ kinase assay.
Results:
Pirfenidone significantly suppressed the expression of CCL18 when the cells were treated with concentrations of 50-250 μg/mL. Pirfenidone did not affect the expression of the IL-4R components. The selective STAT6 inhibitor AS1517499 suppressed CCL18 expression. Both AS1517499 and pirfenidone suppressed STAT6 phosphorylation (p < .05), although the effect of pirfenidone was less marked than that of AS1517499. The Z'-LYTE™ kinase assay showed a reduction in the activities of JAK1, JAK3 and TYK2 by pirfenidone.
Conclusion:
Pirfenidone suppresses CCL18 expression in macrophages and this effect is thought to be attributed partly to the inhibition of STAT6 phosphorylation.
Insights
Pirfenidone, an anti-fibrotic drug, reduces the expression of CC chemokine ligand 18 (CCL18) in macrophages. This effect is partly due to pirfenidone inhibiting STAT6 phosphorylation, a key signaling pathway.
Area of Science:
- Pulmonary medicine
- Immunology
- Pharmacology
Background:
- CC chemokine ligand 18 (CCL18) is implicated in pulmonary fibrosis development.
- Macrophages are a primary source of CCL18.
- The impact of pirfenidone on CCL18 expression in macrophages requires investigation.
Purpose of the Study:
- To determine the effect of pirfenidone on CCL18 expression in macrophages.
- To explore the molecular mechanisms underlying pirfenidone's action on CCL18 production.
Main Methods:
- U937 cells were differentiated into macrophages and stimulated with IL-4 to induce CCL18 production.
- CCL18 mRNA and protein levels were quantified using RT-PCR and ELISA.
- Effects on IL-4 receptor (IL-4R) expression, STAT6 activation, and JAK kinase activity were assessed.
Main Results:
- Pirfenidone significantly suppressed CCL18 expression in a dose-dependent manner (50-250 μg/mL).
- Pirfenidone inhibited STAT6 phosphorylation and reduced JAK1, JAK3, and TYK2 kinase activities.
- Pirfenidone did not alter IL-4R component expression.
Conclusions:
- Pirfenidone effectively suppresses CCL18 expression in macrophages.
- Inhibition of STAT6 phosphorylation contributes to pirfenidone's suppressive effect on CCL18.
- Pirfenidone may modulate JAK-STAT signaling pathways involved in CCL18 production.
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