Evaluation of SK-N-SH Cells as a Model for NMDA Receptor Induced Toxicity.

IF 2.5 Q3 CELL BIOLOGY

Cellular Physiology and Biochemistry Pub Date : 2024-08-30 DOI:10.33594/000000722

Gunnar Goerges, Paul Disse, Stefan Peischard, Nadine Ritter, Christoph Brenker, Guiscard Seebohm, Nathalie Strutz-Seebohm, Julian A Schreiber

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引用次数: 0

Abstract

Background/aims: Over the years, the number of patients with neurodegenerative diseases is constantly rising illustrating the need for new neuroprotective drugs. A promising treatment approach is the reduction of excitotoxicity induced by rising (S)-glutamate levels and subsequent NMDA receptor overactivation. To facilitate the search for new NMDA receptor inhibitors neuronal cell models are needed. In this study, we evaluated the suitability of human SK-N-SH cells to serve as a cell model for neurodegeneration induced by NMDA receptor overstimulation.

Methods: The cytoprotective effect of the unselective NMDA receptor blocker ketamine as well as the GluN2B-selective inhibitor WMS14-10 was evaluated utilizing different cell viability assays, such as endpoint (LDH, CCK-8, DAPI/FACS) and time dependent methods (bioimpedance).

Results: Non-differentiated as well as differentiated SK-N-SH cells express GluN1 and GluN2B subunits. Furthermore, 50 mM (S)-glutamate led to an instantaneous decrease in cell survival. Only application of unselective channel blocker ketamine could protect differentiated cells against this effect, while the selective inhibitor WMS14-10 did not significantly increase cell survival.

Conclusion: SK-N-SH cells show an increased sensitivity to (S)-glutamate mediated cytotoxicity with higher differentiation level, that is only partially induced by NMDA receptor overstimulation. Furthermore, we showed that only unselective NMDA receptor inhibition can partially reverse (S)-glutamate-induced toxicity.

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将 SK-N-SH 细胞作为 NMDA 受体诱导毒性模型进行评估

背景/目的：多年来，神经退行性疾病患者人数不断增加，这说明需要新的神经保护药物。一种很有前景的治疗方法是降低因（S）-谷氨酸水平升高和随后的 NMDA 受体过度激活而诱发的兴奋毒性。为便于寻找新的 NMDA 受体抑制剂，需要建立神经元细胞模型。在这项研究中，我们评估了人 SK-N-SH 细胞作为由 NMDA 受体过度刺激诱导的神经变性细胞模型的适宜性：方法：利用不同的细胞活力检测方法，如终点法（LDH、CCK-8、DAPI/FACS）和时间依赖法（生物阻抗），评估了非选择性NMDA受体阻断剂氯胺酮和GluN2B选择性抑制剂WMS14-10的细胞保护作用：结果：未分化和已分化的 SK-N-SH 细胞均表达 GluN1 和 GluN2B 亚基。此外，50 mM (S)-谷氨酸会导致细胞存活率瞬间下降。只有使用非选择性通道阻断剂氯胺酮才能保护分化细胞免受这种影响，而选择性抑制剂 WMS14-10 并未显著提高细胞存活率：结论：SK-N-SH细胞随着分化水平的提高，对（S）-谷氨酸介导的细胞毒性的敏感性增加，而这种敏感性仅部分由NMDA受体过度刺激诱导。此外，我们还发现，只有非选择性 NMDA 受体抑制才能部分逆转（S）-谷氨酸诱导的毒性。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Cellular Physiology and Biochemistry 生物-生理学

CiteScore

5.80

自引率

0.00%

发文量

审稿时长

1 months

期刊介绍： Cellular Physiology and Biochemistry is a multidisciplinary scientific forum dedicated to advancing the frontiers of basic cellular research. It addresses scientists from both the physiological and biochemical disciplines as well as related fields such as genetics, molecular biology, pathophysiology, pathobiochemistry and cellular toxicology & pharmacology. Original papers and reviews on the mechanisms of intracellular transmission, cellular metabolism, cell growth, differentiation and death, ion channels and carriers, and the maintenance, regulation and disturbances of cell volume are presented. Appearing monthly under peer review, Cellular Physiology and Biochemistry takes an active role in the concerted international effort to unravel the mechanisms of cellular function.