Department of Biomedical and Biotechnological Science, Section of Physiology, University of Catania, Via Santa Sofia, 97-95123 Catania, Italy.
Department of Biomedical and Biotechnological Science, Section of Physiology, University of Catania, Via Santa Sofia, 97-95123 Catania, Italy.
Mol Cell Neurosci. 2018 Apr;88:212-221. doi: 10.1016/j.mcn.2018.02.006. Epub 2018 Feb 8.
The purpose of this research was to explore the behavior of aquaporins (AQPs) in an in vitro model of Parkinson's disease that is a recurrent neurodegenerative disorder caused by the gradual, progressive loss of dopaminergic neurons in the substantia nigra pars compacta. Because of postmortem studies have provided evidences for oxidative damage and alteration of water flow and energy metabolism, we carried out an investigation about AQP4 and 9, demonstrated in the brain to maintain water and energy homeostasis. As an appropriate in vitro cell model, we used SH-SY5Y cultures and induced their differentiation into a mature dopaminergic neuron phenotype with retinoic acid (RA) alone or in association with phorbol-12-myristate-13-acetate (MPA). The association RA plus MPA provided the most complete and mature neuron phenotype, as demonstrated by high levels of β-Tubulin III, MAP-2, and tyrosine hydroxylase. After validation of cell differentiation, the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP) and HO were applied to reproduce a Parkinson's-like stress. The results confirmed RA/MPA differentiated SH-SY5Y as a useful in vitro system for studying neurotoxicity and for using in a MPTP and HO-induced Parkinson's disease cell model. Moreover, the data demonstrated that neuronal differentiation, neurotoxicity, neuroinflammation, and oxidative stress are strongly correlated with dynamic changes of AQP4 and 9 transcription and transduction. New in vitro and in vivo experiments are needed to confirm these innovative outcomes.
本研究旨在探索水通道蛋白（AQP）在帕金森病体外模型中的行为，帕金森病是一种复发性神经退行性疾病，由黑质致密部多巴胺能神经元逐渐进行性丧失引起。由于尸检研究提供了氧化损伤和水流动及能量代谢改变的证据，我们对 AQP4 和 9 进行了研究，AQP4 和 9 在大脑中被证明可维持水和能量内稳态。作为一种合适的体外细胞模型，我们使用 SH-SY5Y 培养物，并单独或与佛波醇-12-肉豆蔻酸-13-乙酸酯（MPA）一起用视黄酸（RA）诱导其分化为成熟的多巴胺能神经元表型。RA 加 MPA 的联合作用提供了最完整和成熟的神经元表型，β-微管蛋白 III、MAP-2 和酪氨酸羟化酶的高水平证明了这一点。在细胞分化得到验证后，应用神经毒素 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）和 HO 来重现帕金森样应激。结果证实，RA/MPA 分化的 SH-SY5Y 可作为研究神经毒性的有用体外系统，并可用于 MPTP 和 HO 诱导的帕金森病细胞模型。此外，数据表明神经元分化、神经毒性、神经炎症和氧化应激与 AQP4 和 9 的转录和转导的动态变化密切相关。需要新的体外和体内实验来证实这些创新结果。
