Mylonas Nikolaos, Siokatas Georgios, Zacharia Effimia, Pol Christine, Rolland Tyler, Kyriazis Ioannis D, Hoffman Matthew, Hildebrand Alycia, Bannister Thomas, Gao Erhe, Goldberg Ira J, Yang Vincent W, Bialkowska Agnieszka B, Elrod John W, Canty John M, Andreadou Ioanna, Weil Brian, Drosatos Konstantinos
Metabolic Biology Laboratory, Cardiovascular Center, Department of Pharmacology, Physiology, and Neurobiology, University of Cincinnati College of Medicine, 231 Albert Sabin Way, CVC-5939, Cincinnati, OH 45267, USA.
Laboratory of Pharmacology, Faculty of Pharmacy, National and Kapodistrian University of Athens, Panepistimioupolis, Zografou, Athens 15771, Greece.
Cardiovasc Res. 2025 Jun 12;121(6):900-914. doi: 10.1093/cvr/cvaf040.
Activation of the transcriptional factor Krüppel-like factor 5 (KLF5) is detrimental to chronic heart failure. We explored the involvement of KLF5 in myocardial ischaemia/reperfusion injury.
Yorkshire pigs underwent 75' of ischaemia, followed by 3 or 24 h of reperfusion. C57BL/6J mice underwent 30' of ischaemia, followed by 10', 2, 12, 24 h, or 4 weeks of reperfusion. Hearts and isolated cardiomyocytes (CMs) were analysed for gene expression. We assessed cardiac function, infarct size (IS), oxidative stress, and fibrosis in mice subjected to pharmacologic or genetic KLF5 inhibition, as well as pharmacologic inhibition of NADPH oxidases or glucose transporter (GLUT)1 and GLUT4. Bulk RNA sequencing, untargeted 1H-NMR metabolomics, and LC-MS lipidomics were performed. Isolated primary murine CMs were infected with recombinant adenovirus expressing KLF5. During reperfusion, CM KLF5 expression was increased in porcine and murine hearts. Pharmacologic or CM-specific genetic inhibition of KLF5 reduced IS and improved cardiac function in mice. Importantly, acute KLF5 inhibition during early reperfusion suppressed fibrosis and preserved systolic cardiac function 4 weeks post-ischaemia/reperfusion. This improvement was associated with lower NADPH-oxidase 4 (NOX4) expression, less oxidative stress, and suppressed inflammation and cell apoptosis. Pharmacologic inhibition of NOX4 conferred the same benefit. Metabolomic analysis indicated that KLF5 inhibition lowered glucose-derived metabolites (UDP-glucose and lactate) at early reperfusion. Accordingly, cardiac GLUT1 and GLUT4 levels were increased with ischaemia/reperfusion, which was reverted by KLF5 inhibition. Pharmacologic inhibition of both GLUT1 and GLUT4 reduced IS. Finally, myocardial KLF5 overexpression increased GLUT1 mRNA levels and mouse mortality.
Ischaemia/reperfusion increases CM KLF5 expression in pigs and mice. This constitutes a central element of myocardial injury pathophysiology and is associated with stimulation of GLUT1 and GLUT4 expression, activation of NOX4, oxidative stress, inflammation, and apoptosis. Acute KLF5 inhibition during reperfusion constitutes a novel therapeutic approach against myocardial ischaemia/reperfusion injury.
转录因子Krüppel样因子5（KLF5）的激活对慢性心力衰竭有害。我们探究了KLF5在心肌缺血/再灌注损伤中的作用。
约克夏猪经历75分钟缺血，随后再灌注3小时或24小时。C57BL/6J小鼠经历30分钟缺血，随后再灌注10分钟、2小时、12小时、24小时或4周。对心脏和分离的心肌细胞（CMs）进行基因表达分析。我们评估了接受药理学或基因KLF5抑制的小鼠的心脏功能、梗死面积（IS）、氧化应激和纤维化，以及NADPH氧化酶或葡萄糖转运蛋白（GLUT）1和GLUT4的药理学抑制情况。进行了批量RNA测序、非靶向1H-NMR代谢组学和LC-MS脂质组学分析。分离的原代小鼠CMs用表达KLF5的重组腺病毒感染。在再灌注期间，猪和小鼠心脏中CM KLF5表达增加。KLF5的药理学或CM特异性基因抑制减少了小鼠的梗死面积并改善了心脏功能。重要的是，早期再灌注期间急性KLF5抑制可抑制纤维化并在缺血/再灌注后4周保留心脏收缩功能。这种改善与较低的NADPH氧化酶4（NOX4）表达、较少的氧化应激以及抑制的炎症和细胞凋亡有关。NOX4的药理学抑制也有同样的益处。代谢组学分析表明，KLF5抑制在再灌注早期降低了葡萄糖衍生的代谢物（UDP-葡萄糖和乳酸）。相应地，缺血/再灌注时心脏GLUT1和GLUT4水平升高，而KLF5抑制可使其恢复。GLUT1和GLUT4的药理学抑制均减少了梗死面积。最后，心肌KLF5过表达增加了GLUT1 mRNA水平并提高了小鼠死亡率。
缺血/再灌注增加了猪和小鼠CMs中KLF5的表达。这构成了心肌损伤病理生理学的核心要素，并与GLUT1和GLUT4表达的刺激、NOX4的激活、氧化应激、炎症和细胞凋亡有关。再灌注期间急性KLF5抑制构成了一种针对心肌缺血/再灌注损伤的新型治疗方法。
