Cell senescence is associated with age-related pathological changes. Increasing evidence has revealed that mitophagy can selectively remove dysfunctional mitochondria. Overexpression of ubiquitin-specific protease 30 (USP30) has been documented to influence mitophagy and deubiquitination of mitochondrial Parkin substrates. This study was conducted to evaluate the roles of USP30 and Parkin in myocardial cell senescence and mitophagy. Initially, myocardial cells were isolated from neonatal SD rats and subjected to d-gal treatment to induce cell senescence, after which the effects of d-gal on mitochondria damage, ROS production, cell senescence, and mitophagy were assessed. The myocardial cells were infected with lentiviruses bearing overexpression plasmids or shRNA targeting Parkin or USP30 to elucidate the effects of Parkin and USP30 on d-gal-induced mitophagy damage and cell senescence. Finally, aging was induced in rats by subcutaneous injection of d-gal to determine the role of Parkin and USP30 on cell senescence in vivo. d-gal was found to trigger mitochondria damage, ROS production, and cell senescence in myocardial cells. The overexpression of Parkin or silencing of USP30 reduced d-gal-induced mitochondrial damage and relieved d-gal-induced myocardial cell senescence. Moreover, the in vivo experiments validated that either elevation of Parkin or silencing USP30 could alleviate d-gal-induced myocardial cell senescence in rats. Silencing USP30 alleviates d-gal-induced mitochondrial damage and consequently suppresses myocardial cell senescence by activating Parkin. Our study highlights the potential of USP30 as a novel target against myocardial cell senescence.

细胞衰老与年龄相关的病理变化有关。越来越多的证据表明，线粒体自噬可以选择性地去除功能失调的线粒体。泛素特异性蛋白酶 30 (USP30) 的过度表达已被证明会影响线粒体 Parkin 底物的线粒体自噬和去泛素化。本研究旨在评估 USP30 和 Parkin 在心肌细胞衰老和线粒体自噬中的作用。最初，从新生 SD 大鼠中分离心肌细胞并进行d -gal 处理以诱导细胞衰老，之后d的作用评估了 -gal 对线粒体损伤、ROS 产生、细胞衰老和线粒体自噬的影响。心肌细胞用带有过表达质粒或靶向 Parkin 或 USP30 的 shRNA 的慢病毒感染，以阐明 Parkin 和 USP30 对d -gal 诱导的线粒体自噬损伤和细胞衰老的影响。最后，通过皮下注射d -gal诱导大鼠衰老，以确定 Parkin 和 USP30 对体内细胞衰老的作用。d -gal 被发现会触发心肌细胞中的线粒体损伤、ROS 产生和细胞衰老。Parkin 的过表达或 USP30 的沉默减少了d -gal 诱导的线粒体损伤并缓解了d-gal 诱导的心肌细胞衰老。此外，体内实验证实，Parkin 的升高或 USP30 的沉默都可以减轻d -gal 诱导的大鼠心肌细胞衰老。沉默 USP30 可减轻d -gal 诱导的线粒体损伤，从而通过激活 Parkin 抑制心肌细胞衰老。我们的研究强调了 USP30 作为抗心肌细胞衰老的新靶点的潜力。