Erastin-sensitive VDAC2 contributes to time-course peritoneal fibrosis via ferroptosis signaling induction.

Abstract

Peritoneal dialysis (PD) is a renal replacement therapy for patients with end-stage renal disease, but long-term treatment often leads to peritoneal fibrosis, compromising dialysis efficacy. This study aimed to elucidate the role of ferroptosis in peritoneal fibrosis (PF) and to clarify the regulatory function of the voltage-dependent anion channel 2 (VDAC2). Mesothelial cell subpopulations were identified from the single-cell dataset GSE130888, followed by differential gene expression analysis to screen ferroptosis-related genes. In vitro and in vivo models were used to validate the involvement of ferroptosis in PF. siRNA-mediated VDAC2 knockdown, combined with the ROS scavenger NAC, was applied to assess its role in regulating mitochondrial ROS during ferroptosis and fibrosis. Single-cell analysis revealed ferroptosis-related gene alterations in both short-term (ST) and long-term (LT) PD, with VDAC2 expression positively associated with dialysis duration. Erastin significantly reduced MET‑5 A cell viability, increased apoptosis, elevated ROS and lipid peroxidation, decreased GSH levels, and upregulated VDAC2, ACSL4, and fibrosis‑related proteins. These effects were reversed by Fer‑1. In vivo, Erastin exacerbated peritoneal dysfunction, collagen deposition, and tissue iron accumulation in mice, and caused complete collapse of cellular architecture, dissolution of organelles, nuclear envelope rupture, and chromatin fragmentation, whereas Fer‑1 improved peritoneal pathology and iron deposition. VDAC2 knockdown restored MET‑5 A cell viability, reduced ROS and lipid peroxidation, increased GSH levels, and downregulated ferroptosis- and fibrosis-related proteins; co‑treatment with NAC further enhanced these protective effects. VDAC2 promotes peritoneal mesothelial cell injury and fibrosis by driving mitochondrial ROS-dependent ferroptosis. Targeting VDAC2 or inhibiting ferroptosis may provide a potential therapeutic strategy to delay peritoneal dialysis-associated fibrosis.