PLGA-loaded puerarin attenuates arsenic-induced liver injury through modulation of ferroptosis, oxidative stress, inflammation, and apoptotic signaling

Arsenic (AR) is an environmental and mining occupational toxicant with reported hepatotoxic effects. Ferroptosis has recently emerged as a critical mechanism involved in arsenic-induced hepatotoxicity. This study investigated the hepatoprotective effects of puerarin (PU) and its PLGA-loaded nanoparticle formulation (PU-PLGA) against arsenic-induced liver injury in rats, with particular emphasis on ferroptosis pathways modulation. Sixty adult male Sprague–Dawley rats were randomly assigned into six groups: control, PU, PU-PLGA, arsenic (AR), AR + PU, and AR + PU-PLGA. Subacute hepatotoxicity was induced by oral administration of arsenic (10 mg/kg) for 14 days. Oxidative stress, ferroptosis-related biomarkers, inflammatory mediators, apoptotic markers, fibrotic indicators, DNA damage, histopathological alterations, and ultrastructural changes were evaluated. Arsenic exposure induced marked hepatic injury characterized by Fe²⁺ accumulation, increased lipid peroxidation markers (MDA and 4-HNE), upregulation of ACSL4, and suppression of GPX4 and SLC7A11, indicating enhanced ferroptotic activity. These alterations were accompanied by oxidative stress, inflammatory activation (NF-κB, TNF-α, IL-6, IL-1β, and COX-2), apoptotic signaling (Bax and caspase-3), DNA damage (8-OHdG and DNA fragmentation), and fibrotic responses (TGF-β and collagen I). Treatment with PU and PU-PLGA significantly attenuated arsenic-induced hepatic injury by reducing iron accumulation and lipid peroxidation, restoring GPX4 and SLC7A11 expression, and suppressing ACSL4 upregulation. Notably, PU-PLGA demonstrated greater efficacy than free PU in modulating ferroptosis-related biomarkers, restoring antioxidant and survival signaling (Nrf2, p-AKT, and Bcl-2), and improving hepatic histopathological and ultrastructural integrity. P PU-PLGA nanoparticles exerted potent hepatoprotective effects against arsenic-induced liver injury, associated with modulation of ferroptosis-related pathways, oxidative stress, inflammatory responses, apoptosis-associated signaling, and profibrotic alterations. These findings highlight PU-PLGA as a promising nanotherapeutic strategy for arsenic-associated hepatotoxicity, although further mechanistic and translational validation is warranted.