Biphasic Effect of Pirfenidone on Angiogenesis

Abstract

Pirfenidone (PFD), a synthetic arsenic compound, has been found to inhibit angiogenesis at high concentrations. However, the biphasic effects of different PFD concentrations on angiogenesis have not yet been elucidated, and the present study used an in vitro model to explore the mechanisms underlying this biphasic response. The effect of PFD on the initial angiogenesis of vascular endothelial cells was investigated through a Matrigel tube formation assay, and the impact of PFD on endothelial cell migration was evaluated through scratch and transwell migration experiments. Moreover, the expression of key migration cytokines, matrix metalloproteinase (MMP)-2 and MMP-9, was examined. Finally, the biphasic mechanism of PFD on angiogenesis was explored through cell signaling and apoptosis analyses. The results showed that 10–100 μM PFD has a significant and dose-dependent inhibitory effect on tube formation and migration, while 10 nM–1 μM PFD significantly promoted tube formation and migration, with 100 nM PFD having the strongest effect. Additionally, we found that a high concentration of PFD could significantly inhibit MMP-2 and MMP-9 expression, while low concentrations of PFD significantly promoted their expression. Finally, we found that high concentrations of PFD inhibited EA.hy926 cell tube formation by promoting apoptosis, while low concentrations of PFD promoted tube formation by increasing MMP-2 and MMP-9 protein expression predominantly via the EGFR/p-p38 pathway. Overall, PFD elicits a biphasic effect on angiogenesis through different mechanisms, could be used as a new potential drug for the treatment of vascular diseases.

A Protective Effect of Pirfenidone in Lung Fibroblast–Endothelial Cell Network via Inhibition of Rho-Kinase Activity

“…To date, the influence of pirfenidone on the vascular endothelium has not been sufficiently examined; however, pirfenidone has been suggested to act on the vascular endothelium in a biphasic fashion [ 22 , 23 ]. Pirfenidone has been shown to promote angiogenesis at low concentrations and to inhibit neoangiogenesis at high concentrations [ 23 ].…”

“…To date, the influence of pirfenidone on the vascular endothelium has not been sufficiently examined; however, pirfenidone has been suggested to act on the vascular endothelium in a biphasic fashion [ 22 , 23 ]. Pirfenidone has been shown to promote angiogenesis at low concentrations and to inhibit neoangiogenesis at high concentrations [ 23 ]. High concentrations of pirfenidone significantly inhibit the expression of key migration cytokines, matrix metalloproteinases (MMPs) MMP-2 and MMP-9; by contrast, low concentrations of pirfenidone significantly promote expression of these substances [ 23 ].…”

“…Pirfenidone has been shown to promote angiogenesis at low concentrations and to inhibit neoangiogenesis at high concentrations [ 23 ]. High concentrations of pirfenidone significantly inhibit the expression of key migration cytokines, matrix metalloproteinases (MMPs) MMP-2 and MMP-9; by contrast, low concentrations of pirfenidone significantly promote expression of these substances [ 23 ]. High concentrations of pirfenidone have also been reported in previous study to promote apoptosis [ 23 ].…”

“…High concentrations of pirfenidone significantly inhibit the expression of key migration cytokines, matrix metalloproteinases (MMPs) MMP-2 and MMP-9; by contrast, low concentrations of pirfenidone significantly promote expression of these substances [ 23 ]. High concentrations of pirfenidone have also been reported in previous study to promote apoptosis [ 23 ]. In that study, high concentrations of pirfenidone were defined as 10–100 μM, which differs from the concentrations used in our experimental system.…”

“…The literature contains limited reports describing how antifibrotic agents used in clinical practice affect the vascular network in interstitial pneumonia. Specifically, pirfenidone has been reported to reduce vascular endothelial network formation in vascular endothelial cells, and the inhibitory effect of pirfenidone on tube formation has been studied [ 22 , 23 ]. As these reports were based on studies of pirfenidone in vascular endothelial cells alone, further investigation is necessary to evaluate the real-life pharmacological effects of this agent on the vascular network in interstitial pneumonia.…”