Abstract

Objective Atherosclerosis (AS) is a chronic inflammatory disease characterized by lipid deposition. Ferroptosis is a form of cell death driven by the accumulation of peroxidized lipids. N 4 -acetylcytidine (ac 4 C) is an RNA modification catalyzed by N-acetyltransferase 10 (NAT10). This study aimed to elucidate the effect and mechanism of NAT10-mediated ac 4 C modification on ferroptosis in an in vitro cellular model of AS. Methods Human vascular endothelial cells (HVECs) were treated with oxidized low-density lipoprotein (ox-LDL) to establish a cell model. A dot blot assay was used to assess the ac 4 C level, and quantitative real-time polymerase chain reaction (qPCR) and Western blot were used to detect NAT10 expression in ox-LDL-treated HVECs. The function of NAT10 was evaluated in vitro using a cell counting kit-8, lactate dehydrogenase, Fe 2+ , malondialdehyde, reactive oxygen species, and glutathione detection kits, as well as PI staining. The underlying mechanism was investigated using ac 4 C RNA immunoprecipitation (RIP), bioinformatic analysis, a dual-luciferase reporter assay, and RNA stability assays. Results The levels of ac 4 C and NAT10 were elevated in ox-LDL-stimulated HVECs. Knockdown of NAT10-inhibited ferroptosis in ox-LDL-treated HVECs. Furthermore, NAT10 knockdown reduced the ac 4 C level and mRNA stability of TFRC. Additional analysis revealed that TFRC overexpression reversed the inhibitory effect on ferroptosis in HVECs following NAT10 knockdown. Conclusions In conclusion, we demonstrated that silencing of NAT10-inhibited ferroptosis in HVECs by reducing the ac 4 C modification and mRNA stability of TFRC. These findings suggest a potential novel mechanism relevant to AS pathogenesis, although further in vivo validation is warranted.