Department of Ophthalmology, Gavin Herbert Eye Institute, University of California Irvine, Irvine, CA 92697, USA.
Department of Pathology and Laboratory Medicine, University of California Irvine, Irvine, CA 92697, USA.
Aging (Albany NY). 2020 May 29;12(10):9031-9040. doi: 10.18632/aging.103164.
Citicoline is the exogenous form of the nootropic, Cytidine 5'-diphosphate-choline that exerts its neuroprotective effects in the brain as well as in the eye. The current study characterized the cytoprotective effects of purified Citicoline in transmitochondrial AMD (Age-related Macular Degeneration) RPE cybrid cells which carry diseased mitochondria from clinically characterized AMD patients. The effects of Citicoline were examined via flow cytometry analysis of AnnexinV/ PI-stained cells, IncuCyte live-cell imaging analysis to quantify cells undergoing caspase-3/7-mediated apoptosis, analyses of gene expression profiles of apoptosis, hypoxia, and angiogenesis markers, and measurement of ROS levels and cell viability. Our results demonstrated that Citicoline when added exogenously alleviates apoptotic effects as evidenced by diminished AnnexinV/PI and Caspase-3/7 staining, downregulation of apoptosis genes, enhanced cell viability, and reduced oxidative stress in AMD RPE cybrid cells. In conclusion, our study identified Citicoline as a protector in AMD RPE cybrid cells . However, further studies are required to establish the merit of Citicoline as a cytoprotective molecule in AMD and to decipher the molecular underpinnings of its mechanism of action in AMD.
胞磷胆碱是一种神经营养药物,外源性胞磷胆碱(Cytidine 5'-diphosphate-choline)在大脑和眼睛中发挥其神经保护作用。本研究对携带临床特征性 AMD 患者疾病线粒体的传代线粒体 AMD(Age-related Macular Degeneration)RPE 细胞杂种进行了纯化胞磷胆碱的细胞保护作用的鉴定。通过 AnnexinV/PI 染色细胞的流式细胞术分析、定量 caspase-3/7 介导的细胞凋亡的 IncuCyte 活细胞成像分析、细胞凋亡、缺氧和血管生成标志物的基因表达谱分析以及 ROS 水平和细胞活力的测量,检测了胞磷胆碱的作用。结果表明,胞磷胆碱通过减少 AnnexinV/PI 和 Caspase-3/7 染色、下调凋亡基因、增强细胞活力和降低 AMD RPE 细胞的氧化应激,减轻了细胞凋亡作用。总之,本研究确定胞磷胆碱是 AMD RPE 细胞杂种的保护剂。然而,还需要进一步的研究来确定胞磷胆碱作为 AMD 细胞保护分子的价值,并阐明其在 AMD 中的作用机制的分子基础。