YAP 通过促进 ACSL4 的 NEDD4L 介导的泛素化和降解来减轻心肌缺血再灌注损伤中的铁死亡。
YAP Facilitates NEDD4L-Mediated Ubiquitination and Degradation of ACSL4 to Alleviate Ferroptosis in Myocardial Ischemia-Reperfusion Injury.
机构信息
Cardiovascular Surgery ICU, Second Xiangya Hospital of Hunan Province, Changsha, Hunan Province, China.
Cardiopulmonary Bypass, Second Xiangya Hospital of Hunan Province, Changsha, Hunan Province, China.
出版信息
Can J Cardiol. 2023 Nov;39(11):1712-1727. doi: 10.1016/j.cjca.2023.07.030. Epub 2023 Aug 2.
BACKGROUND
Ferroptosis is a novel iron-dependent type of cell death that takes part in the progression of myocardial ischemia/reperfusion injury (MIRI). However, the detailed mechanism of ferroptosis underlying MIRI remains unclear. This study aimed to investigate the regulatory role of yes-associated protein (YAP) in ferroptosis during MIRI.
METHODS
The in vivo and in vitro MIRI models were established in the Sprague-Dawley (SD) rats and H9C2 cardiomyocytes. The infarct volume, pathologic changes, cardiac function, serum levels of lactate dehydrogenase (LDH) and creatine kinase (CK)-MB were detected. Western blotting and immunohistochemistry were performed to measure the expression of YAP, neural precursor cell expressed developmentally downregulated 4-like (NEDD4L) and ferroptosis-related proteins. Ferroptosis was evaluated by Fe, malondialdehyde (MDA), LDH, glutathione (GSH), and lipid reactive oxygen species (ROS) levels. Molecular mechanism was analyzed by co-immunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP), and dual-luciferase reporter assay.
RESULTS
YAP and NEDD4L were remarkably low expressed in MIRI models. YAP overexpression reduced myocardial infarct volume and improved cardiac function. In addition, YAP inhibited MIRI-induced ferroptosis as confirmed by reducing levels of Fe, MDA, LDH, lipid ROS, and ferroptosis-related protein ACSL4, and enhancing GSH level and cell viability. Mechanistically, YAP facilitated NEDD4L transcription that consequently caused ubiquitination and degradation of ACSL4, thereby restraining ferroptosis in MIRI. YAP knockdown aggravated MIRI-induced ferroptosis, which was counteracted by NEDD4L overexpression.
CONCLUSIONS
YAP represses MIRI-induced cardiomyocyte ferroptosis via promoting NEDD4L transcription and subsequent ubiquitination and degradation of ACSL4. YAP-mediated ferroptosis inhibition might be a novel therapeutic strategy for MIRI.
背景
铁死亡是一种新的铁依赖性细胞死亡方式，参与心肌缺血/再灌注损伤（MIRI）的进展。然而，MIRI 中铁死亡的详细机制尚不清楚。本研究旨在探讨 YAP 在 MIRI 中铁死亡中的调节作用。
方法
在 Sprague-Dawley（SD）大鼠和 H9C2 心肌细胞中建立体内和体外 MIRI 模型。检测梗死面积、病理变化、心功能、血清乳酸脱氢酶（LDH）和肌酸激酶同工酶（CK-MB）水平。Western blot 和免疫组化检测 YAP、神经前体细胞表达发育下调 4 样（NEDD4L）和铁死亡相关蛋白的表达。通过铁、丙二醛（MDA）、LDH、谷胱甘肽（GSH）和脂质活性氧（ROS）水平评估铁死亡。通过共免疫沉淀（Co-IP）、染色质免疫沉淀（ChIP）和双荧光素酶报告基因分析分析分子机制。
结果
YAP 和 NEDD4L 在 MIRI 模型中表达明显降低。YAP 过表达可减少心肌梗死面积，改善心功能。此外，YAP 抑制 MIRI 诱导的铁死亡，表现为降低铁、MDA、LDH、脂质 ROS 和铁死亡相关蛋白 ACSL4 的水平，增加 GSH 水平和细胞活力。机制上，YAP 促进 NEDD4L 的转录，导致 ACSL4 的泛素化和降解，从而抑制 MIRI 中的铁死亡。YAP 敲低加重 MIRI 诱导的铁死亡，而 NEDD4L 的过表达可逆转这一现象。
结论
YAP 通过促进 NEDD4L 转录和随后的 ACSL4 泛素化和降解来抑制 MIRI 诱导的心肌细胞铁死亡。YAP 介导的铁死亡抑制可能是 MIRI 的一种新的治疗策略。
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