Protective effect of mitochondria‑targeted peptide MTP‑131 against oxidative stress‑induced apoptosis in RGC‑5 cells.
Protective effect of mitochondria‑targeted peptide MTP‑131 against oxidative stress‑induced apoptosis in RGC‑5 cells.
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DOI：
10.3892/mmr.2017.6271
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发表时间：
2017-04
影响因子：
3.4
通讯作者：
Wang K
中科院分区：
文献类型：
--
作者：
Chen M;Liu B;Ma J;Ge J;Wang K
人眼的视网膜极易受到氧化损伤。研究表明，氧化应激是老年性黄斑变性、糖尿病性视网膜病变、青光眼和视网膜色素变性发病的主要机制。MTP-131是一种新型的靶向于线粒体的多肽，在体外和动物模型中被证明特异性地集中在线粒体内膜中，并表现出显著的抗氧化作用。在本研究中，在视网膜神经节细胞系RGC-5中评估MTP-131对过氧化氢（H2 O2）诱导的氧化损伤的保护作用。通过乳酸脱氢酶（LDH）测定来测量细胞活力。流式细胞仪检测线粒体膜电位的变化，激光共聚焦显微镜检测细胞内活性氧（ROS）的产生。Annexin V-异硫氰酸荧光素/碘化丙啶染色用于评估细胞凋亡。通过共聚焦显微镜分析细胞色素c的释放。与单独用H2 O2处理的细胞相比，用MTP-131预处理的细胞以剂量依赖性方式抑制H2 O2诱导的细胞毒性并减少LDH释放。线粒体去极化和ROS的产生也被MTP-131预处理阻止。此外，MTP-131预处理抑制细胞色素c从线粒体释放到细胞质，并显着减少RGC-5细胞的凋亡，与单独用H2 O2处理的细胞相比。结论：MTP-131对氧化应激诱导的RGC-5细胞凋亡具有保护作用，为年龄相关性视网膜疾病的治疗提供了新的途径。
The retina of the human eye is extremely vulnerable to oxidative damage. Previous studies have demonstrated that oxidative stress is the predominant mechanism associated with the pathogenesis of age-related macular degeneration, diabetic retinopathy, glaucoma and retinitis pigmentosa. MTP-131, a novel mitochondria-targeted peptide, has been demonstrated to specifically concentrate in the inner mitochondria membrane and to exhibit remarkable antioxidant effects both in vitro and in animal models. In the present study, the protective effect of MTP-131 was evaluated in response to hydrogen peroxide (H2O2)-induced oxidative damage in a retinal ganglion cell line, RGC-5. Cell viability was measured by lactate dehydrogenase (LDH) assay. Changes of mitochondrial membrane potential and generation of intracellular reactive oxygen species (ROS) were measured by flow cytometry and confocal microscopy, respectively. Annexin V-fluorescein isothiocyanate/propidium iodide staining was used for assessment of apoptosis. Release of cytochrome c was analyzed by confocal microscopy. Pretreatment of cells with MTP-131 inhibited H2O2-induced cytotoxicity and reduced LDH release in a dose-dependent manner, compared with cells treated with H2O2 alone. Mitochondrial depolarization and ROS generation were also prevented by MTP-131 pretreatment. In addition, MTP-131 pretreatment inhibited cytochrome c release from mitochondria to cytoplasm, and significantly reduced apoptosis in RGC-5 cells, compared with cells treated with H2O2 alone. In conclusion, mitochondria-targeted peptide MTP-131 exhibited a protective effect against oxidative stress-induced apoptosis in RGC-5 cells, which may provide a novel approach for the treatment of age-associated retinal diseases.