ALKBH5 ALLEVIATES HYPOXIA POSTCONDITIONING INJURY IN d-GALACTOSE–INDUCED SENESCENT CARDIOMYOCYTES BY REGULATING STAT3
ALKBH5 ALLEVIATES HYPOXIA POSTCONDITIONING INJURY IN d-GALACTOSE–INDUCED SENESCENT CARDIOMYOCYTES BY REGULATING STAT3
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ALKBH 5通过调节STAT 3减轻d-半乳糖诱导的衰老心肌细胞缺氧后损伤
DOI：
10.1097/shk.0000000000002031
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发表时间：
2023-01
期刊：
影响因子：
3.1
通讯作者：
Xuan Zhang;Feixiang Li;Ji Ma;Xiaobei Zhang;Yiqing Yin
中科院分区：
文献类型：
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作者：
Xuan Zhang;Feixiang Li;Ji Ma;Xiaobei Zhang;Yiqing Yin
摘要缺血后处理（I/Post）通过激活内源性心肌保护机制，如JAK/信号转导子和转录激活子3（STAT 3）和PI 3 K/Akt通路，减轻I/R损伤，这是传统的心肌保护方法。根据先前的研究，I/Post的心脏保护作用可能在老年小鼠中丧失，并且在我们先前的研究中，缺氧后处理（H/Post）在衰老的心肌细胞中缺乏保护作用，这与长非编码RNA H19的低表达有关。N6-甲基腺苷（m6 A）修饰是一个动态和可逆的过程，已被证实在心血管疾病中发挥作用。然而，心肌I/Post中m6 A修饰的机制仍有待探讨。从2日龄Sprague-Dawley大鼠分离新生心肌细胞，并通过d-半乳糖诱导衰老，随后通过缺氧-复氧和H/Post刺激。通过细胞活力和Bcl-2/Bax蛋白比值评价缺氧损伤。使用比色m6 A RNA甲基化定量试剂盒测量总m6 A水平，并通过MeRIP（甲基化RNA免疫沉淀）确定m6 A修饰和差异表达的mRNA。我们发现H/Post增加了老年心肌细胞中m6 A甲基化并降低了RNA mA脱甲基酶alkB同源物5（ALKBH 5）的表达。此外，ALKBH 5敲低加剧了衰老心肌细胞H/Post后的损伤。此外，ALKBH 5通过介导其m6 A修饰和长非编码RNA H19/miR-124- 3 p来调节STAT 3的表达。ALKBH 5还减轻了衰老心肌细胞中STAT 3低表达诱导的H/Post损伤。
ABSTRACT Ischemic postconditioning (I/Post) reduces I/R injury by activating endogenous cardioprotection mechanisms, such as the JAK/signal transducer and activator of transcription 3 (STAT3) and PI3K/Akt pathways, which offer a traditional approach to myocardial protection. According to a previous study, cardioprotection by I/Post may be lost in aged mice, and in our previous research, hypoxic postconditioning (H/Post) lacked a protective effect in senescent cardiomyocytes, which was associated with low expression of long noncoding RNA H19. The N6-methyladenosine (m6A) modification is a dynamic and reversible process that has been confirmed to play a role in cardiovascular diseases. However, the mechanisms of m6A modification in myocardial I/Post remain to be explored. Neonatal cardiomyocytes were isolated from 2-day-old Sprague-Dawley rats, and senescence was induced by d-galactose, followed by stimulation of hypoxia-reoxygenation and H/Post. Hypoxic injury was evaluated by cell viability and the Bcl-2/Bax protein ratio. Total m6A levels were measured using a colorimetric m6A RNA Methylation Quantification Kit, and the m6A modified and differentially expressed mRNA was determined by MeRIP (methylated RNA immunoprecipitation). We found that H/Post increased m6A methylation and decreased RNA mA demethylase alkB homolog 5 (ALKBH5) expression in aged cardiomyocytes. Furthermore, ALKBH5 knockdown exacerbated injury following H/Post in senescent cardiomyocytes. In addition, ALKBH5 regulated STAT3 expression by mediating its m6A modification and long noncoding RNA H19/miR-124-3p. ALKBH5 also alleviated the H/Post injury induced by the low expression of STAT3 in senescent cardiomyocytes.