Department of Ophthalmology, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey.
Neuroscience Research Center, Suleyman Demirel University, Isparta, Turkey.
Graefes Arch Clin Exp Ophthalmol. 2021 Jun;259(6):1539-1554. doi: 10.1007/s00417-021-05074-7. Epub 2021 Feb 5.
Bevacizumab (BEV) is a blocker of circulating VEGF A generation. However, BEV has adverse apoptotic and cytotoxic effects via upregulation of mitochondrial reactive oxygen species (ROS) and TRPM2 activation, and downregulation of cytosolic glutathione (GSH) in neuronal cells. We investigated the possible protective effects of GSH treatment on BEV-induced oxidant and apoptotic adverse actions in the TRPM2 expressing adult retinal pigment epithelial-19 (ARPE-19) and SH-SY5Y neuronal cells.
The ARPE-19 and SH-SY5Y cells were divided into five main groups: Control, GSH (10 mM for 2 h), BEV (0.25 mg/ml for 24 h), BEV+GSH, and BEV+TRPM2 channel blockers (ACA or 2-APB). In the SH-SY5Y cells, the Ca analyses (Fluo-3) were performed only, although Fluo-3 and the remaining analyses were performed in the ARPE-19 cells.
The levels of apoptosis, cell death, mitochondrial ROS, lipid peroxidation, caspase-3, caspase-9, ADP-ribose-induced TRPM2 current density, cytosolic-free Zn, and Ca were increased by BEV, although their levels were diminished by the treatments of GSH and TRPM2 blockers. The BEV-induced decreases of cell viability, GSH levels, and glutathione peroxidase activities were increased by the treatment of GSH. BEV-induced increase of TRPM2 expression was decreased by the treatment of GSH, although BEV-induced decrease of VEGF A expression was further decreased by the treatment of GSH.
Our data confirmed that BEV-induced mitochondrial ROS and apoptosis in the human retinal epithelial cells were modulated by GSH and TRPM2 inhibition. The treatment of GSH may be considered as a therapeutic approach to BEV-induced ARPE-19 cell injury.
贝伐单抗（BEV）是一种阻断循环 VEGF A 生成的药物。然而，BEV 通过上调线粒体活性氧（ROS）和 TRPM2 的激活，以及下调神经元细胞中的细胞溶质谷胱甘肽（GSH），产生不良的凋亡和细胞毒性作用。我们研究了 GSH 处理对 BEV 诱导的成年视网膜色素上皮-19（ARPE-19）和 SH-SY5Y 神经元细胞中氧化应激和凋亡不良作用的可能保护作用。
将 ARPE-19 和 SH-SY5Y 细胞分为五个主要组：对照组、GSH（10 mM，2 h）、BEV（0.25 mg/ml，24 h）、BEV+GSH 和 BEV+TRPM2 通道阻滞剂（ACA 或 2-APB）。在 SH-SY5Y 细胞中，仅进行了 Ca 分析（Fluo-3），而在 ARPE-19 细胞中进行了 Fluo-3 和其余分析。
BEV 增加了细胞凋亡、细胞死亡、线粒体 ROS、脂质过氧化、caspase-3、caspase-9、ADP-核糖诱导的 TRPM2 电流密度、细胞溶质游离 Zn 和 Ca 的水平，而 GSH 和 TRPM2 阻滞剂的处理则降低了这些水平。GSH 处理增加了 BEV 诱导的细胞活力、GSH 水平和谷胱甘肽过氧化物酶活性的降低，而 GSH 处理增加了 BEV 诱导的 TRPM2 表达的增加，尽管 GSH 处理进一步降低了 BEV 诱导的 VEGF A 表达的降低。
我们的数据证实，GSH 和 TRPM2 抑制调节了 BEV 诱导的人视网膜上皮细胞中线粒体 ROS 和凋亡。GSH 的治疗可能被认为是治疗 BEV 诱导的 ARPE-19 细胞损伤的一种方法。
