Ma Xiaofeng, Chen Wang, Hu Zhihao, Xie Like, Li Zhaobing, Liu Huan, Li Zili, Jiang Zhentao, Huang Jiangwei, Jiang Changrong, Huang Ke, Xiao Sujun
The Affiliated Nanhua Hospital, Department of Cardiology, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China.
The Affiliated Nanhua Hospital, Department of Cardiology, Hengyang Medical School, University of South China, Hengyang, Hunan, 421001, China.
Exp Cell Res. 2025 Jul 1;450(1):114613. doi: 10.1016/j.yexcr.2025.114613. Epub 2025 May 28.
The anthracycline chemotherapeutic agent doxorubicin (DOX) is widely used to treat cancer. However, DOX carries a high risk of adverse effects such as cardiovascular events. The aim of this study was to explore the role of Trim65 in DOX-induced cardiotoxicity (DIC) and its underlying mechanisms.
Cellular studies were performed by exposing H9c2 cells (rat cardiomyocytes) to DOX. H9c2 cells were infected with lentivirus encoding negative control (LV-NC) or Trim65 (LV-Trim65). C57BL/6J mice were exposed to adeno-associated virus 9 (AAV9) containing cTnT promoter-encoded Trim65 sequence (AAV-Trim65) or AAV9 negative control sequence (AAV-NC) via tail vein injection. Furthermore, the following analysis were performed: cell viability, intracellular ROS production and lipid peroxidation. Echocardiography was used to measure the heart function of the mice. qPCR and western blotting were used to assess the expression of Trim65, p53, SLC7A11, and GPX4. According to our study, Trim65 expression was significantly downregulated in DIC. Overexpression of Trim65 exhibited considerable protection against to DIC, confirmed by both in vitro and in vivo experiments. In DOX-treated mice, mitochondria deformation was observed in the heart, as well as a high level of lipid peroxidation (signs of ferroptosis) and iron content, which were mitigated by overexpression of Trim65. Mechanistically, our study confirmed that DOX reduced the Trim65-mediated ubiquitination of p53, ultimately inhibiting the degration of p53. We also found that Trim65 mitigated DOX-induced ferroptosis via p53.
Collectively, our data demonstrated that Trim65 inhibits ferroptosis by degrading p53, thereby alleviating DIC. Therefore, Trim65 could be a promising target for the treatment of DIC.
蒽环类化疗药物阿霉素（DOX）被广泛用于治疗癌症。然而，DOX具有较高的不良反应风险，如心血管事件。本研究的目的是探讨Trim65在阿霉素诱导的心脏毒性（DIC）中的作用及其潜在机制。
通过将H9c2细胞（大鼠心肌细胞）暴露于DOX进行细胞研究。H9c2细胞用编码阴性对照（LV-NC）或Trim65（LV-Trim65）的慢病毒感染。C57BL/6J小鼠通过尾静脉注射暴露于含有心肌肌钙蛋白T启动子编码的Trim65序列的腺相关病毒9（AAV9）（AAV-Trim65）或AAV9阴性对照序列（AAV-NC）。此外，还进行了以下分析：细胞活力、细胞内活性氧生成和脂质过氧化。超声心动图用于测量小鼠的心脏功能。qPCR和蛋白质印迹法用于评估Trim65、p53、溶质载体家族7成员11（SLC7A11）和谷胱甘肽过氧化物酶4（GPX4）的表达。根据我们的研究，Trim65表达在DIC中显著下调。体外和体内实验均证实，Trim65的过表达对DIC具有显著的保护作用。在DOX处理的小鼠中，观察到心脏中的线粒体变形，以及高水平的脂质过氧化（铁死亡迹象）和铁含量，而Trim65的过表达减轻了这些现象。从机制上讲，我们的研究证实DOX降低了Trim65介导的p53泛素化，最终抑制了p53的降解。我们还发现Trim65通过p53减轻了DOX诱导的铁死亡。
总体而言，我们的数据表明Trim65通过降解p53抑制铁死亡，从而减轻DIC。因此，Trim65可能是治疗DIC的一个有前景的靶点。
