Abstract
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 in vitro. 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.
Highlights
Citicoline is the international nonproprietary name given to the exogenous pharmacological form of Cytidine 5'-diphosphate-choline (CDP-Choline, CDPCho), a naturally occurring endogenous nucleotide compound that is water-soluble and has a molecular weight of 488.32 g/mol [1, 2]
Our current study identified the cytoprotective potential of exogenously added purified Citicoline in transmitochondrial AMD RPE cybrid cells in vitro
Using a combination of apoptotic assays, we found that Citicoline mitigates apoptotic cell death as evidenced by diminished Annexin V/ propidium iodide (PI) positive cell population, reduced Caspase-3/7-mediated apoptosis in live cells, downregulation of apoptotic genes, and enhanced cell viability in Citicoline-treated transmitochondrial AMD RPE cybrid cells
Summary
Citicoline is the international nonproprietary name given to the exogenous pharmacological form of Cytidine 5'-diphosphate-choline (CDP-Choline, CDPCho), a naturally occurring endogenous nucleotide compound that is water-soluble and has a molecular weight of 488.32 g/mol [1, 2]. By activating the biosynthesis of structural phospholipids, Citicoline maintains neuronal membrane integrity, influences neurotransmitter levels, increases norepinephrine and dopamine levels in the central nervous system, restores the activity of membrane sodium/potassium ATPase and mitochondrial ATPase, and enhances brain function [1]. Owing to these mechanisms, Citicoline has been successfully used as a neuroprotective agent to prevent neuronal aging and improve memory and learning in vivo [4]. Parisi et al demonstrated that Citicoline injected intramuscularly improves retinal and visual function in glaucoma patients [9]
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