{"title":"ALKBH5 Regulates Inflammation and Pyroptosis of Coronary Heart Disease by Targeting SPEN in a YTHDF1-Mediated Manner.","authors":"Shan Huang, Jizhang Huang, Yongguang Lu, Zewen Hong, Guoyong Lai, Yunyu Chen","doi":"10.1007/s10557-025-07746-6","DOIUrl":null,"url":null,"abstract":"<p><strong>Purpose: </strong>Coronary heart disease (CHD) occurs when the arteries supplying blood to the heart become narrowed or blocked owing to plaque buildup, leading to reduced blood flow and potential heart attacks. N6-methyladenosine (m<sup>6</sup>A) modification is a common form of post-transcriptional RNA modification. ALKB homolog 5 (ALKBH5) is an RNA demethylase that specifically removes the m<sup>6</sup>A modification from RNA. This study aimed to investigate the role of ALKBH5 in CHD and the underlying mechanism.</p><p><strong>Methods: </strong>Both cellular and animal CHD models were established. The expression levels of Alkbh5 and fibrosis-related markers were analyzed by qRT-PCR. Cell viability and cytotoxicity were assessed via cell counting kit-8. Inflammatory cytokines levels were detected by ELISA. Flow cytometry was used to detect pyroptosis. The interaction between ALKBH5/YTH N6-methyladenosine RNA binding protein (YTHDF)1 and SPEN transcriptional repressor (SPEN) was examined through RNA immunoprecipitation and dual-luciferase reporter assays.</p><p><strong>Results: </strong>ALKBH5-mediated demethylation of m<sup>6</sup>A was decreased in CHD rat heart tissues and oxidized low-density lipoprotein (ox-LDL)-treated H9c2 cells. In addition, Alkbh5 overexpression increased the cell viability and suppressed the inflammation, pyroptosis, and fibrosis in ox-LDL-treated H9c2 cells. In in vivo studies, Alkbh5 overexpression reduced myocardial injury and fibrosis in CHD rats by suppressing inflammation and pyroptosis. Mechanically, Alkbh5 overexpression decreased the stability of Spen mRNA. Additionally, ALKBH5/YTHDF1 m<sup>6</sup>A axis regulated the expression of Spen. Moreover, Ythdf1 overexpression counteracted ALKBH5-mediated inhibition of inflammation, pyroptosis, and fibrosis in ox-LDL-treated H9c2 cells.</p><p><strong>Conclusion: </strong>ALKBH5 regulated inflammation and pyroptosis of CHD by targeting SPEN in a YTHDF1-mediated manner, which could provide a reference for CHD treatment.</p>","PeriodicalId":9557,"journal":{"name":"Cardiovascular Drugs and Therapy","volume":" ","pages":""},"PeriodicalIF":3.1000,"publicationDate":"2025-07-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cardiovascular Drugs and Therapy","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1007/s10557-025-07746-6","RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"CARDIAC & CARDIOVASCULAR SYSTEMS","Score":null,"Total":0}
引用次数: 0
Abstract
Purpose: Coronary heart disease (CHD) occurs when the arteries supplying blood to the heart become narrowed or blocked owing to plaque buildup, leading to reduced blood flow and potential heart attacks. N6-methyladenosine (m6A) modification is a common form of post-transcriptional RNA modification. ALKB homolog 5 (ALKBH5) is an RNA demethylase that specifically removes the m6A modification from RNA. This study aimed to investigate the role of ALKBH5 in CHD and the underlying mechanism.
Methods: Both cellular and animal CHD models were established. The expression levels of Alkbh5 and fibrosis-related markers were analyzed by qRT-PCR. Cell viability and cytotoxicity were assessed via cell counting kit-8. Inflammatory cytokines levels were detected by ELISA. Flow cytometry was used to detect pyroptosis. The interaction between ALKBH5/YTH N6-methyladenosine RNA binding protein (YTHDF)1 and SPEN transcriptional repressor (SPEN) was examined through RNA immunoprecipitation and dual-luciferase reporter assays.
Results: ALKBH5-mediated demethylation of m6A was decreased in CHD rat heart tissues and oxidized low-density lipoprotein (ox-LDL)-treated H9c2 cells. In addition, Alkbh5 overexpression increased the cell viability and suppressed the inflammation, pyroptosis, and fibrosis in ox-LDL-treated H9c2 cells. In in vivo studies, Alkbh5 overexpression reduced myocardial injury and fibrosis in CHD rats by suppressing inflammation and pyroptosis. Mechanically, Alkbh5 overexpression decreased the stability of Spen mRNA. Additionally, ALKBH5/YTHDF1 m6A axis regulated the expression of Spen. Moreover, Ythdf1 overexpression counteracted ALKBH5-mediated inhibition of inflammation, pyroptosis, and fibrosis in ox-LDL-treated H9c2 cells.
Conclusion: ALKBH5 regulated inflammation and pyroptosis of CHD by targeting SPEN in a YTHDF1-mediated manner, which could provide a reference for CHD treatment.
期刊介绍：
Designed to objectively cover the process of bench to bedside development of cardiovascular drug, device and cell therapy, and to bring you the information you need most in a timely and useful format, Cardiovascular Drugs and Therapy takes a fresh and energetic look at advances in this dynamic field.
Homing in on the most exciting work being done on new therapeutic agents, Cardiovascular Drugs and Therapy focusses on developments in atherosclerosis, hyperlipidemia, diabetes, ischemic syndromes and arrhythmias. The Journal is an authoritative source of current and relevant information that is indispensable for basic and clinical investigators aiming for novel, breakthrough research as well as for cardiologists seeking to best serve their patients.
Providing you with a single, concise reference tool acknowledged to be among the finest in the world, Cardiovascular Drugs and Therapy is listed in Web of Science and PubMed/Medline among other abstracting and indexing services. The regular articles and frequent special topical issues equip you with an up-to-date source defined by the need for accurate information on an ever-evolving field. Cardiovascular Drugs and Therapy is a careful and accurate guide through the maze of new products and therapies which furnishes you with the details on cardiovascular pharmacology that you will refer to time and time again.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
