N6-Methyladenosine Demethylase ALKBH5 Promotes Pyroptosis by Modulating PTBP1 mRNA Stability in LPS-Induced Myocardial Dysfunction.

IF 1.8 4区医学 Q3 CARDIAC & CARDIOVASCULAR SYSTEMS Acta Cardiologica Sinica Pub Date : 2024-05-01 DOI:10.6515/ACS.202405_40(3).20240127A

Min Liu, Xiyun Chen

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Abstract

Objective: This study aims to investigate the mechanism by which alkB homolog 5 (ALKBH5) regulates polypyrimidine tract-binding protein 1 (PTBP1) to mediate cardiomyocyte pyroptosis in sepsis-induced myocardial injury.

Methods: Lipopolysaccharide (LPS)-exposed H9C2 cell and rat models were established to mimic septic myocardial injury both in vitro and in vivo. The mRNA and protein levels of ALKBH5 and PTBP1 in the LPS-induced cell and septic rat models were detected. CCK-8 and flow cytometry were applied to detect cell viability and pyroptosis. H&E staining was used to observe myocardial tissue damage in rats, and immunohistochemistry to analyze the expression of pyroptosis and inflammation-related proteins in rat tissues.

Results: Elevated expressions of both ALKBH5 and PTBP1 were found in the myocardial tissues of LPS-induced septic rats. ALKBH5 knockdown could restore the cell viability and cell pyroptosis inhibited by LPS, while ALKBH5 promoted PTBP1 mRNA stability by affecting its N6-methyladenosine (m6A) modification. In vivo experiments showed that PTBP1 knockdown could largely reverse the antiproliferative and pro-pyroptosis effects of ALKBH5 in LPS-exposed H9C2 cells. ALKBH5 knockdown in in vivo experiments was found to suppress the expressions of pyroptosis biomarkers and attenuate myocardial injury in septic rats.

Conclusions: ALKBH5 promoted mRNA stability and the expression of PTBP1 through m6A modification to induce pyroptosis in cardiomyocytes and ultimately aggravate sepsis-induced myocardial dysfunction.

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N6-甲基腺苷脱甲基酶 ALKBH5 通过调节 LPS 诱导的心肌功能障碍中 PTBP1 mRNA 的稳定性促进脓毒症的发生

研究目的本研究旨在探讨烷基同源物 5（ALKBH5）调控多嘧啶束结合蛋白 1（PTBP1）在脓毒症诱发的心肌损伤中介导心肌细胞脓毒症的机制：方法：建立了暴露于脂多糖（LPS）的 H9C2 细胞和大鼠模型，在体外和体内模拟脓毒症心肌损伤。检测LPS诱导的细胞和败血症大鼠模型中ALKBH5和PTBP1的mRNA和蛋白水平。应用 CCK-8 和流式细胞术检测细胞存活率和脓毒症。用 H&E 染色法观察大鼠心肌组织损伤，用免疫组化法分析大鼠组织中热休克和炎症相关蛋白的表达：结果：LPS诱导的脓毒症大鼠心肌组织中ALKBH5和PTBP1的表达均升高。敲除 ALKBH5 可恢复 LPS 抑制的细胞活力和细胞裂解，而 ALKBH5 则通过影响 N6-甲基腺苷（m6A）修饰促进 PTBP1 mRNA 的稳定性。体内实验表明，在暴露于 LPS 的 H9C2 细胞中，敲除 PTBP1 可在很大程度上逆转 ALKBH5 的抗增殖和促嗜突变作用。在体内实验中发现，ALKBH5敲除可抑制脓毒症大鼠体内脓毒症生物标志物的表达，减轻心肌损伤：结论：ALKBH5通过m6A修饰促进mRNA的稳定性和PTBP1的表达，诱导心肌细胞的脓毒症，最终加重脓毒症诱发的心肌功能障碍。

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来源期刊

Acta Cardiologica Sinica 医学-心血管系统

CiteScore

2.90

自引率

15.80%

发文量

144

审稿时长

>12 weeks

期刊介绍： Acta Cardiologica Sinica welcomes all the papers in the fields related to cardiovascular medicine including basic research, vascular biology, clinical pharmacology, clinical trial, critical care medicine, coronary artery disease, interventional cardiology, arrythmia and electrophysiology, atherosclerosis, hypertension, cardiomyopathy and heart failure, valvular and structure cardiac disease, pediatric cardiology, cardiovascular surgery, and so on. We received papers from more than 20 countries and areas of the world. Currently, 40% of the papers were submitted to Acta Cardiologica Sinica from Taiwan, 20% from China, and 20% from the other countries and areas in the world. The acceptance rate for publication was around 50% in general.