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Alkylation Repair Homolog 5 Regulates N(6)-methyladenosine (m6A) Methylation of Mitsugumin 53 to Attenuate Myocardial
Dong Li1, Lianggang Li, Shiyong Dong
1Department of Cardiovascular Surgery, The First Medical Center of Chinese PLA General Hospital, Beijing, China.
Abstract:
N(6)-methyladenosine (m6A) methylation modification is involved in the progression of myocardial infarction (MI). In this study, we investigated the effects of demethylase alkylation repair homolog 5 (ALKBH5) on cell apoptosis and oxidative stress in MI. The ischemia/reperfusion (I/R) injury mouse model and hypoxia/reoxygenation (H/R) cell model were established. The levels of ALKBH5 and mitsugumin 53 (MG53) were measured by quantitative real-time polymerase chain reaction, immunohistochemical, and immunofluorescence analysis. Apoptosis was evaluated by TUNEL assay, flow cytometry, and western blot. Oxidative stress was assessed by antioxidant index kits. Methylation was analyzed by RNA binding protein immunoprecipitation (RIP), MeRIP, and dual-luciferase reporter assay. We observed that ALKBH5 and MG53 were highly expressed in MI. Overexpression of ALKBH5 inhibited H/R-induced cardiomyocyte apoptosis and oxidative stress in vitro, and it inhibited I/R-induced collagen deposition, cardiac function, and apoptosis in vivo. ALKBH5 could bind to MG53, inhibit m6A methylation of MG53, and increase its mRNA stability. Silencing of MG53 counteracted the inhibition of apoptosis and oxidative stress induced by ALKBH5. In conclusion, ALKBH5 suppressed m6A methylation of MG53 and inhibited MG53 degradation to inhibit apoptosis and oxidative stress of cardiomyocytes, thereby attenuating MI. The results provided a theoretical basis that ALKBH5 is a potential target for MI treatment.
Insights
Demethylase alkylation repair homolog 5 (ALKBH5) protects against myocardial infarction (MI) by inhibiting cell apoptosis and oxidative stress. ALKBH5 targets mitsugumin 53 (MG53) to stabilize its mRNA, thereby reducing MI progression.
Area of Science:
- Biochemistry
- Molecular Biology
- Cardiovascular Research
Background:
- N(6)-methyladenosine (m6A) methylation is implicated in myocardial infarction (MI) progression.
- The role of the demethylase alkylation repair homolog 5 (ALKBH5) in MI pathogenesis requires further investigation.
Purpose of the Study:
- To investigate the effects of ALKBH5 on cardiomyocyte apoptosis and oxidative stress in the context of MI.
- To elucidate the molecular mechanism by which ALKBH5 influences MI progression.
Main Methods:
- Establishment of ischemia/reperfusion (I/R) mouse and hypoxia/reoxygenation (H/R) cell models.
- Quantitative real-time PCR, immunohistochemistry, immunofluorescence, TUNEL assay, flow cytometry, western blot, and RIP-MeRIP assays were employed.
- Analysis of ALKBH5, mitsugumin 53 (MG53) expression, apoptosis, oxidative stress, and m6A methylation levels.
Main Results:
- ALKBH5 and MG53 were upregulated in MI models.
- ALKBH5 overexpression attenuated H/R-induced apoptosis and oxidative stress in vitro and I/R-induced cardiac dysfunction and apoptosis in vivo.
- ALKBH5 bound to MG53, reduced its m6A methylation, and enhanced mRNA stability, with MG53 silencing counteracting ALKBH5's protective effects.
Conclusions:
- ALKBH5 suppresses m6A methylation of MG53, enhancing its stability and inhibiting cardiomyocyte apoptosis and oxidative stress.
- ALKBH5 plays a protective role in MI by modulating the MG53 pathway.
- ALKBH5 presents a potential therapeutic target for treating myocardial infarction.
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