This study investigates the role of N-acetyltransferase 10 (NAT10) in cardiomyocyte ferroptosis during ischemia-reperfusion (I/R) injury. NAT10 was found to be upregulated in mouse hearts and cardiomyocytes exposed to I/R, and this upregulation was transcriptionally activated by p53. Overexpression of NAT10 in cardiomyocytes exacerbated ferroptosis and I/R injury, while knocking down NAT10 had the opposite effects. Mechanistically, NAT10 induced ac4C modification of Mybbp1a, increasing its stability and activating p53, which subsequently repressed the transcription of the anti-ferroptotic gene SLC7A11. Knockdown of Mybbp1a partially abrogated the detrimental effects of NAT10 overexpression on ferroptosis and I/R injury. These findings suggest that the NAT10/Mybbp1a/p53 axis forms a positive feedback loop that promotes cardiomyocyte ferroptosis and I/R injury, providing a novel therapeutic target for cardiac I/R injury.This study investigates the role of N-acetyltransferase 10 (NAT10) in cardiomyocyte ferroptosis during ischemia-reperfusion (I/R) injury. NAT10 was found to be upregulated in mouse hearts and cardiomyocytes exposed to I/R, and this upregulation was transcriptionally activated by p53. Overexpression of NAT10 in cardiomyocytes exacerbated ferroptosis and I/R injury, while knocking down NAT10 had the opposite effects. Mechanistically, NAT10 induced ac4C modification of Mybbp1a, increasing its stability and activating p53, which subsequently repressed the transcription of the anti-ferroptotic gene SLC7A11. Knockdown of Mybbp1a partially abrogated the detrimental effects of NAT10 overexpression on ferroptosis and I/R injury. These findings suggest that the NAT10/Mybbp1a/p53 axis forms a positive feedback loop that promotes cardiomyocyte ferroptosis and I/R injury, providing a novel therapeutic target for cardiac I/R injury.