Abstract—One of the leading mechanisms of neurodegeneration in Parkinson’s disease is excitotoxicity of glutamate, the main excitatory brain neurotransmitter. Excitotoxicity develops as a result of excessive stimulation of ionotropic NMDA glutamate receptors. Drugs reversibly inhibiting NMDA receptors are considered as promising pharmacological agents that prevent the death of neurons and slow down neurodegeneration. In this work, we studied the effect of N-(2-adamantyl)-hexamethyleneimine hydrochloride (hemantane) on changes in the intracellular concentration of Ca²⁺ ([Ca²⁺]_i), Na⁺ ([Na⁺]_i) and mitochondrial potential (ΔΨ_m) induced by NMDA in cultured neurons. Cultures were prepared from the cerebral cortex of 1-2-day-old Wistar rats. Experiments with animals were performed in accordance with the ethical principles and regulatory documents recommended by the European Convention on the Protection of Vertebrate Animals. The measurements of [Ca²⁺]_i, [Na⁺]_i and ΔΨ_m were taken using a fluorescence microscopy system based on an Olympus IX-71 inverted microscope with a Sutter Lambda 10-2 multi-wavelength illumination system and a cooled CoolSnap HQ CCD camera. The system was controlled using the MetaFluor software. It was shown that short-term subtoxic doses of NMDA (2–3 min, 10 μM) cause a rapid rise in [Ca²⁺]_i, which is reversibly inhibited by high concentrations of Mg²⁺ (10 mM) and hemantane (100 μM). Prolonged administration of the neurotoxic NMDA concentration (20 min, 500 μM) caused the development of delayed calcium deregulation (DCD), a steady rise in [Na⁺]_i and a profound drop in ΔΨ_m. Hemantane (100 μM) reduced [Ca²⁺]_i elevation, postponed the development of DCD, reduced mitochondrial depolarization, and helped to restore the initial values of [Ca²⁺]_i, [Na⁺]_i and ΔΨ_m after NMDA washout. Unlike hemantane, the high-affinity non-competitive inhibitor of NMDA channels MK-801 irreversibly blocked changes in [Ca²⁺]_i and ΔΨ_m, even at a high NMDA concentration (500 μM). Obviously, hemantane exhibits neuroprotective properties due to a decrease in Ca²⁺ and Na⁺ fluxes through NMDA channels and a lower elevation of [Ca²⁺]_i and [Na⁺]_i. The reversibility of NMDA channel inhibition favors the normalization of brain functioning after termination of hemantane administration.

摘要—帕金森氏病的神经退行性疾病的主要机制之一是谷氨酸的兴奋性毒性，谷氨酸是一种主要的兴奋性脑神经递质。过度刺激离子型NMDA谷氨酸受体会导致兴奋性中毒。可逆地抑制NMDA受体的药物被认为是有前途的药理药物，可防止神经元死亡并减缓神经变性。在这项工作中，我们研究了N-（2-金刚烷基）-六亚甲基亚胺盐酸盐（金刚烷）对Ca 2 ⁺（[Ca ²⁺ ] _i），Na ⁺（[Na ⁺ ] _i）细胞内浓度变化的影响。和线粒体电位（ΔΨ_米NMDA在培养的神经元中诱导）。从1-2天大的Wistar大鼠的大脑皮层制备培养物。根据《欧洲保护脊椎动物动物公约》所建议的道德原则和法规文件对动物进行了实验。[Ca的测量²⁺ ]_我，的[Na ⁺ ]_我和ΔΨ_米用荧光显微镜系统基于采取与萨特LAMBDA 10-2多波长照明系统的Olympus IX-71倒置显微镜和冷却CoolSnap HQ CCD相机。使用MetaFluor软件控制系统。结果表明，NMDA的短期亚毒性剂量（2-3分钟，10μM）会导致[Ca ²⁺] _i，可被高浓度的Mg ²⁺（10 mM）和金刚烷（100μM）可逆地抑制。神经毒性NMDA浓度（20分钟，500μM）的延长给药引起的延迟的钙失调的（DCD），稳步上升中的[Na发展⁺ ]_我和ΔΨ一个深刻降_米。Hemantane（100μM）还原的[Ca ²⁺ ]_我抬高，推迟DCD的发展，降低了线粒体去极化，并帮助恢复[Ca的初始值²⁺ ]_我，的[Na ⁺ ]_我和ΔΨ_米NMDA冲洗后。不像hemantane，NMDA通道MK-801的高亲和力的非竞争性抑制剂不可逆地阻断的[Ca变化²⁺ ]_我和ΔΨ_米，即使在高浓度的NMDA（500μM）。显然，由于通过NMDA通道的Ca ²⁺和Na ^+的通量减少以及[Ca ²⁺ ] _i和[Na ⁺ ] _i的升高较低，金丝丹具有神经保护作用。NMDA通道抑制的可逆性有利于终止金刚烷给药后脑功能的正常化。