The in vitro anti-fibrotic effect of Pirfenidone on human pterygium fibroblasts is associated with down-regulation of autocrine TGF-β and MMP-1

Abstract

We aimed to investigate the in vitro effect of pirfenidone (PFD) on proliferation, migration and collagen contraction of human pterygium fibroblasts (HPFs). HPFs were obtained from tissue explants during pterygium surgery. After treatment with pirfenidone, the HPFs proliferation was measured by MTT, cell cycle progression measured by flow cytometry, cell migration measured by the scratch assay, and cell contractility evaluated in fibroblast-populated collagen gels. The expression of TGF-β1, TGF-β2, MMP-1 and TIMP-1 were also determined with quantitative PCR, western blot and immunofluorescence staining. Results showed pirfenidone markedly inhibited HPFs proliferation with an IC50 of approximately 0.2 mg/ml. After treatment with 0.2 mg/ml pirfenidone for 24 hours, HPFs were at G0/G1 cell cycle arrest, with significantly reduced cell migration capability and collagen contraction, decreased mRNA and protein expressions of TGF-β1, TGF-β2 and MMP-1, and no alterations of TIMP-1 expression. Thus, we have concluded that pirfenidone at 0.2 mg/ml inhibits proliferation, migration, and collagen contraction of HPFs, which is associated with decreased expression of TGF-β and MMP-1, and pirfenidone might represent a potentially therapeutic agent to prevent the recurrence of pterygium after surgery.

Pirfenidone attenuates synovial fibrosis and postpones the progression of osteoarthritis by anti-fibrotic and anti-inflammatory properties in vivo and in vitro

“…The precise mechanisms of action of PFD remain undefined, although it is approved by the FDA for the treatment of idiopathic pulmonary fibrosis [ 33 ]. Previous studies have confirmed that PFD decreases autocrine expression of TGF-β and MMP-1 in human pterygium fibroblasts [ 34 ] and inhibits the production of collagen I in human intestinal fibroblasts[ 31 ]. PFD decreases the deposition of hydroxyapatite by osteoblasts in a dose-dependent manner, which may inhibit osteophyte formation [ 35 ].…”

Effect of Pirfenidone on TGF-β1-Induced Myofibroblast Differentiation and Extracellular Matrix Homeostasis of Human Orbital Fibroblasts in Graves’ Ophthalmopathy

“…Pirfenidone is an anti-inflammatory and antifibrotic agent employed for the treatment of fibrotic processes in several animal models and the affected cells in the lung, liver, kidney, and heart [11][12][13][14][15][16]. Studies have confirmed the effect of pirfenidone on the inhibition of myofibroblast differentiation, fibrotic protein and profibrotic cytokine release, and ECM deposition [17][18][19][20][21][22]. Although it is evident that pirfenidone prevents fibrosis by reducing the reaction induced by TGF-β1, the definite signaling pathways remain to be determined.…”

“…In vitro and in vivo studies have confirmed its effect on the inhibition of myofibroblast differentiation, proliferation, and profibrotic cytokine release in lung, liver, heart, and renal tissues [12][13][14][15][16]. Throughout these years, pirfenidone has been used in several experimental disease models including renal and hepatic fibrosis [17], systemic sclerosis [18], wound healing [19], and ocular pathologies [20][21][22]. In terms of ocular disease, pirfenidone has been reported to play an antifibrotic and anti-inflammatory role in vitro, particularly in the orbital fibroblasts [20], pterygium fibroblasts [21], and retinal pigment epithelial cells [22].…”

“…Throughout these years, pirfenidone has been used in several experimental disease models including renal and hepatic fibrosis [17], systemic sclerosis [18], wound healing [19], and ocular pathologies [20][21][22]. In terms of ocular disease, pirfenidone has been reported to play an antifibrotic and anti-inflammatory role in vitro, particularly in the orbital fibroblasts [20], pterygium fibroblasts [21], and retinal pigment epithelial cells [22]. However, its molecular targets and actions in GO have not been elucidated.…”

Expert Perspectives On Clinical Challenges: Expert Perspectives: Challenges in Scleroderma

“…Pirfenidone. Pirfenidone is an antifibrotic agent approved by the FDA for the treatment of IPF, and although its mechanism of action is not completely understood, it appears to target transforming growth factor β signaling, a key feature in the pathogenesis of fibrosis (67,68). An open-label phase II study of pirfenidone (69) showed an acceptable tolerability profile, especially with a longer titration schedule, despite its well-recognized gastrointestinal side effect profile.…”