Pirfenidone Inhibits Hypoxic Pulmonary Hypertension through the NADPH/ROS/p38 Pathway in Adventitial Fibroblasts in the Pulmonary Artery

Abstract

Hypoxic pulmonary hypertension (HPH) is a devastating disease characterized by progressive vasoconstriction and vascular remodeling. Pirfenidone (PFD) inhibits the progression of HPH, though the molecular mechanisms remain unknown. This study is aimed at determining the role and mechanism of PFD in HPH in human pulmonary artery adventitial fibroblasts (HPAAFs), which were cultured under normal or hypoxic conditions. NOX4 and Rac1 were inhibited or overexpressed by shRNA or pcDNA3.1, respectively. Proliferation of HPAAFs was quantified by colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) assays to assess cellular metabolic activity, cell counts, and ethynyldeoxyuridine (EdU) assays to detect DNA synthesis. Migration of HPAAFs was assessed by a wound healing assay. The expression levels of smooth muscle alpha-actin (a-SMA) and procollagen I (COL1A1) were assessed by RT-PCR and western blot analysis. PFD suppressed hypoxia-induced proliferation and migration of HPAAFs. Compared with the hypoxic control group, PFD reduced the expression of a-SMA and procollagen I (COL1A1). PFD reduced hypoxia-induced phosphorylation of p38 through the NOX4/reactive oxygen species (ROS) signaling pathway. Moreover, Rac1 also decreased hypoxia-induced phosphorylation of p38, without any cross-interaction with NOX4. These findings demonstrate that PFD is a novel therapeutic agent to prevent cell proliferation, migration, and fibrosis, which might be useful in inhibiting vascular remodeling in patients with HPH.

Cancer-Associated Fibroblasts in the Hypoxic Tumor Microenvironment

“…Pirfenidone, an anti-fibrotic agent for idiopathic pulmonary fibrosis, inhibits CAF activation and proliferation in lung and pancreatic cancers by targeting TGF-β and reduces the production of fibrotic mediators secreted by CAFs [ 114 , 115 , 116 , 117 ]. In hypoxic pulmonary hypertension, pirfenidone inhibits hypoxia-induced proliferation and migration of adventitial fibroblasts, thereby reducing the expression of α-SMA and collagen type I alpha 1 chain (COL1A1) [ 118 ]. Minnelide, a pro-drug of triptolide, downregulates TGF-β signaling in CAFs to enhance tumor regression in a mouse model of pancreatic cancer.…”

Metabolic Dysregulation in Idiopathic Pulmonary Fibrosis

“…Indeed, hypoxia induces the proliferation and differentiation of pulmonary artery adventitial fibroblasts into myofibroblasts that ultimately, stimulate the recruitment of inflammatory cells and the release of key regulators. The proliferation and differentiation of pulmonary artery adventitial fibroblasts during chronic hypoxia are mainly dependent on the signal transduction of the p38 pathway and its downstream mediator, HIF-1 [ 66 ].…”

CeO2/Bovine Serum Albumin Nanoclusters with Robust Reactive Oxygen Species Scavenging and Improved Endothelial Dysfunction Abilities for the Treatment of Pulmonary Hypertension

“…However, cck8 (Figure b) and EdU staining (Figure c,d) results showed that CeO 2 /BSA nanoclusters inhibited the proliferation of PMECs induced by hypoxia, which may be attributed to its ability of scavenging ROS. Overmigration and overproliferation coexist and influence each other in PMEC dysfunction . Hence, we also monitored the migration of PMECs treated with CeO 2 /BSA nanoclusters using a scratch test.…”