Zhang Yuanyuan, Yang Yang, Xie Zhen, Zuo Wenqi, Jiang Hongyan, Zhao Xueyan, Sun Yu, Kong Weijia
Department of Otorhinolaryngology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 1277 Jiefang Avenue, Wuhan, 430022, China.
Department of Otolaryngology-Head and Neck Surgery, Renmin Hospital, Wuhan University, 238 Jiefang Road, Wuhan, 430060, China.
Mol Neurobiol. 2016 Nov;53(9):5971-5984. doi: 10.1007/s12035-015-9469-7. Epub 2015 Nov 2.
Oxidative damage to the inner ear is responsible for several types of sensorineural deafness. Cochlear stria marginal cells (MCs) are thought to be vulnerable to such oxidative stress. Activated poly(ADP-ribose) polymerase 1 (PARP1) has been implicated in several diseases, but the effect of PARP1 on MCs subjected to oxidative stress remains elusive. In this study, we established an in vitro cellular oxidative stress model using glucose oxidase (GO) and attempted to explore the role that PARP1 plays in the oxidative damage of MCs. In this study, PARP1 and poly-ADP-ribose (PAR) were highly expressed in GO-treated MCs, and this was accompanied by loss of MC viability, excessive generation of reactive oxygen species (ROS), collapse of mitochondria membrane potential (ΔΨm), and redistribution of the mitochondrial downstream pathway-related molecules Bax and cytochrome c, eventually causing MC death. These effects were almost completely counteracted by suppressing PARP1 expression with small interfering RNA (siRNA). We also found that caspase-3 activation was a downstream event of PARP activation and that apoptosis of MCs was suppressed, although not completely, by pretreatment with the pan-caspase inhibitor z-VAD-fmk. The suppression was less than that when PARP1 expression was inhibited. We conclude that GO treatment induces activation of PARP1, which causes MC damage via mitochondrial mediation. PARP1 plays a pivotal role in GO-induced MC death, at least in part, via the caspase-3 cascade. Our study might provide a new cellular and molecular approach for the treatment of oxidative stress-related sensorineural deafness.
内耳的氧化损伤是多种类型感音神经性耳聋的病因。耳蜗边缘细胞(MCs)被认为易受这种氧化应激的影响。活化的聚(ADP - 核糖)聚合酶1(PARP1)与多种疾病有关,但PARP1对遭受氧化应激的MCs的影响仍不清楚。在本研究中,我们使用葡萄糖氧化酶(GO)建立了体外细胞氧化应激模型,并试图探究PARP1在MCs氧化损伤中所起的作用。在本研究中,PARP1和聚 - ADP - 核糖(PAR)在GO处理的MCs中高表达,同时伴随着MCs活力丧失、活性氧(ROS)过度生成、线粒体膜电位(ΔΨm)崩溃以及线粒体下游途径相关分子Bax和细胞色素c的重新分布,最终导致MCs死亡。用小干扰RNA(siRNA)抑制PARP1表达几乎完全抵消了这些影响。我们还发现caspase - 3激活是PARP激活的下游事件,并且用泛caspase抑制剂z - VAD - fmk预处理可抑制MCs凋亡,尽管不完全抑制。这种抑制作用小于抑制PARP1表达时的作用。我们得出结论,GO处理诱导PARP1激活,PARP1通过线粒体介导导致MCs损伤。PARP1至少部分地通过caspase - 3级联反应在GO诱导的MCs死亡中起关键作用。我们的研究可能为治疗氧化应激相关的感音神经性耳聋提供一种新的细胞和分子方法。