m6A levels were significantly elevated in the hypoxia/reoxygenation cell model.
Reduced expression of Alkbh5 mRNA was observed alongside elevated m6A levels in the hypoxia/reoxygenation model.
Overexpression of Alkbh5 inhibited that increased in the hypoxia/reoxygenation model.
Alkbh5 overexpression decreased m6A levels of Ythdf1 and promoted Fth1 translation by enhancing Ythdf1 mRNA expression.
Knockdown of Ythdf1 restored ferroptosis in the hypoxia/reoxygenation model, counteracting the effects of Alkbh5 overexpression.
In a myocardial ischemia/reperfusion injury rat model, Alkbh5 overexpression alleviated myocardial injury and increased Fth1 protein levels.
Simplified
BACKGROUND: Myocardial infarction (MI) is a leading cause of global mortality. , an iron-dependent form of programmed cell death, has recently emerged as a critical player in cardiovascular diseases. N6-methyladenosine (m6A), the most prevalent RNA methylation modification in eukaryotic cells, has been implicated in various pathological processes; however, its regulatory role in MI through ferroptosis remains poorly understood. This study aimed to elucidate the mechanism by which mediates MI via ferroptosis.
METHODS: A hypoxia/reoxygenation (H/R) model was established using H9C2 cells to simulate myocardial injury. RNA methylation levels were quantified via dot blot assay. Ferroptosis was evaluated by measuring lactate dehydrogenase (LDH) release, Felevels, glutathione (GSH), lipid reactive oxygen species (ROS), malondialdehyde (MDA), and apoptosis. The underlying molecular mechanisms were investigated using western blotting, quantitative real-time PCR (qPCR), methylated RNA immunoprecipitation (MeRIP), and RIP. Findings were further validated in a myocardial ischemia/reperfusion injury (MIRI) rat model. 2+
RESULTS: The results revealed that m6A levels were significantly elevated in the H/R cell model, accompanied by reduced expression of Alkbh5 mRNA. Moreover, Alkbh5 overexpression inhibited ferroptosis increased in the H/R model. Mechanistically, Alkbh5 overexpression decreased m6A levels of Ythdf1 and H9C2 cells while promoting Fth1 translation by enhancing Ythdf1 mRNA expression. Knockdown of Ythdf1 restored ferroptosis in the H/R model, counteracting the effects of Alkbh5 overexpression. Furthermore, Alkbh5 overexpression alleviated myocardial injury in the MIRI rat model, upregulated Ythdf1 mRNA expression, and increased Fth1 protein levels.
CONCLUSION: This study demonstrates that Alkbh5 ameliorates MI by inhibiting ferroptosis through m6A demethylation of Fth1. These findings provide novel insights into the molecular mechanisms underlying MI and highlight potential therapeutic targets for MI treatment.
Key numbers
m6A levels increased
Increase in m6A levels
Compared to control group in H/R models
LDH release decreased
Decrease in LDH release
Measured in H9C2 cells
GSH levels increased
Increase in GSH levels
Observed in H9C2 cells after Alkbh5 overexpression
Full Text
We can’t show the full text here under this license.
Declarations. Ethics approval and consent to participate: This study was approved by the Ethics Committee of MDKN Biotechnology Co., Lt (Approval No. MDKN-2024-048). All animal experiments should comply with the ARRIVE guidelines. All methods were carried out in accordance with relevant guidelines and regulations. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests. Clinical trial number: Not applicable.
