Astaxanthin ameliorates perfluorooctane sulfonate-induced liver lipid metabolism disorders by targeting ACSL4-mediated ferroptosis

<i>Background</i>: Perfluorooctane sulfonate (PFOS), a persistent organic pollutant, induces hepatotoxicity by disrupting hepatic lipid metabolism, with effective therapeutic interventions currently limited. Astaxanthin (ASX), a potent antioxidant carotenoid, exhibits various health benefits. This study aimed to investigate the hepatoprotective effects of ASX against PFOS-induced liver injury and elucidate the underlying mechanisms. <i>Methods</i>: <i>In vivo</i>, C57BL/6J mice were pretreated with ASX (25, 50, or 100 mg per kg per bw) or ferrostatin-1 (Fer-1; 1 mg per kg per bw), followed by PFOS exposure (10 mg per kg per bw) for 28 days. <i>In vitro</i>, AML12 cells were pretreated with 40 μM ASX or 1 μM Fer-1 before 200 μM PFOS treatment. RNA-sequence analysis was conducted to explore the possible targets for the action of ASX and further validated it by establishing a stable acyl-CoA synthetase long-chain family member 4 (ACSL4) knockout hepatocyte cell line. <i>Result</i>: Transcriptome profiling identified ferroptosis as a potential pathway in PFOS-mediated hepatotoxicity. ASX supplementation significantly attenuated PFOS-induced hepatic lipid deposition and hepatocellular injury in cell and animal models. RNA sequencing revealed that ASX alleviated PFOS-induced hepatic injury by modulating ferroptosis, with ACSL4 identified as a potential target. <i>In vivo</i> and <i>in vitro</i> experiments further demonstrated that ASX significantly suppressed PFOS-induced hepatocyte ferroptosis, while ACSL4 knockout weakened this protective effect on liver cells. <i>Conclusions</i>: Our study suggests that ASX ameliorates PFOS-induced liver injury by targeting ACSL4-mediated ferroptosis. Dietary ASX supplementation may represent a promising therapeutic strategy for preventing PFOS-associated liver diseases.