Pirfenidone inhibits epidural scar fibroblast proliferation and differentiation by regulating TGF-β1-induced Smad-dependent and -independent pathways.

Epidural fibrosis causes serious complications in patients who have undergone laminectomy. Pirfenidone is an effective antifibrotic agent but its effect on epidural fibrosis remains unclear. In this study, we aimed to investigate the effect of pirfenidone on epidural fibrosis and to evaluate its mechanism of action on human epidural scar fibroblasts. In a rat model of laminectomy, the degree of epidural fibrosis was quantified via Rydell standard classification, histological analysis, and collagen density analyses. In cultured human epidural scar fibroblasts, cell proliferation was measured using a Cell Counting Kit-8 and EdU assay. Cell apoptosis was detected using Annexin V/propidium iodide staining, and cytotoxicity was evaluated via lactate dehydrogenase assay. Relative mRNA levels of α-smooth muscle actin (α-SMA) and collagen type I were analyzed using quantitative polymerase chain reaction. The protein expression of α-SMA and collagen type I and the phosphorylation status of Smad2, Smad3, protein kinase B (Akt), and p38 were determined via western blotting. Pirfenidone reduced epidural fibrosis by inhibiting fibroblast proliferation and suppressing collagen formation in rats. It also inhibited human epidural scar fibroblast proliferation with no cytotoxic or apoptotic effects. Pirfenidone inhibited fibroblast differentiation by decreasing TGF-β1-induced transcriptional and translational expression of α-SMA. It inhibited TGF-β1-induced phosphorylation of Smad2, Smad3, Akt, and p38. This study suggests that topical application of pirfenidone could reduce epidural scar adhesion after laminectomy, and that its mechanism of action may be the inhibition of TGF-β1-induced epidural scar fibroblast proliferation and differentiation into myofibroblasts through the attenuation of TGF-β1-induced Smad-dependent and -independent pathways.