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Pirfenidone inhibits transforming growth factor β1-induced extracellular matrix production in nasal polyp-derived
Jae-Min Shin1, Joo-Hoo Park, Il-Ho Park
1Department of Otorhinolaryngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea.
Purpose:
Pirfenidone has been shown to have antifibrotic and anti-inflammatory effects in the lungs. The purpose of this study was to evaluate the inhibitory effects of pirfenidone on transforming growth factor (TGF)-β1-induced myofibroblast differentiation and extracellular matrix accumulation. We also determined the molecular mechanisms of pirfenidone in nasal polyp-derived fibroblasts (NPDF).
Methods:
NPDFs were isolated from nasal polyps from eight patients who had chronic rhinosinusitis with nasal polyp. Pirfenidone was used to treat TGF-β1-induced NPDFs. Cytotoxicity was evaluated by using a 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-tetrazolium bromide assay. Fibroblast migration was evaluated with scratch assays. Expression levels of α-smooth muscle actin (SMA), fibronectin, and phosphorylated Smad2/3 were determined by Western blot and/or reverse transcription-polymerase chain reaction and immunofluorescent staining. Total collagen production was analyzed with the Sircol collagen assay and contractile activity was measured by a collagen gel contraction assay.
Results:
Pirfenidone (0-2 mg/mL) has no significant cytotoxic effects in TGF-β1-induced NPDFs. Migration of NPDFs was significantly inhibited by pirfenidone treatment. The expression levels of α-SMA and fibronectin were significantly reduced in pirfenidone-treated NPDFs. Collagen contraction and production were also significantly decreased by pirfenidone treatment. Finally, pirfenidone significantly inhibited phosphorylation of the Smad2/3 pathway in TGF-β1-induced NPDFs.
Conclusions:
Pirfenidone has an inhibitory effect on TGF-β1-induced migration, myofibroblast differentiation (α-SMA), extracellular matrix accumulation, and collagen contraction by blocking the phosphorylation of Smad2/3 pathways in NPDFs. Thus, pirfenidone may inhibit TGF-β1-induced extracellular matrix by regulating Smad2/3.
Insights
Pirfenidone inhibits nasal polyp fibroblast migration, differentiation, and extracellular matrix accumulation. This antifibrotic effect is mediated by blocking transforming growth factor-beta1 (TGF-β1) signaling pathways.
Area of Science:
- Fibrosis research
- Cell biology
- Pharmacology
Background:
- Pirfenidone exhibits antifibrotic and anti-inflammatory properties.
- Transforming growth factor-beta1 (TGF-β1) is a key mediator of fibrosis.
- Nasal polyp-derived fibroblasts (NPDFs) are relevant models for studying fibrotic processes.
Purpose of the Study:
- To investigate pirfenidone's inhibitory effects on TGF-β1-induced myofibroblast differentiation.
- To assess pirfenidone's impact on extracellular matrix accumulation in NPDFs.
- To elucidate the molecular mechanisms underlying pirfenidone's action in NPDFs.
Main Methods:
- NPDFs were isolated and treated with pirfenidone and TGF-β1.
- Assays included cytotoxicity (MTT), migration (scratch), protein expression (Western blot, immunofluorescence), and collagen production/contraction.
- Smad2/3 pathway phosphorylation was analyzed.
Main Results:
- Pirfenidone demonstrated no cytotoxicity in NPDFs.
- Significant inhibition of NPDF migration, α-smooth muscle actin (α-SMA) and fibronectin expression was observed.
- Pirfenidone reduced collagen production and contraction, and inhibited Smad2/3 phosphorylation.
Conclusions:
- Pirfenidone effectively inhibits TGF-β1-induced fibroblast responses in nasal polyps.
- The drug's antifibrotic effects are linked to the downregulation of the Smad2/3 signaling pathway.
- Pirfenidone holds potential for treating fibrotic conditions in the upper airway.
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