Arzhanov Ivan, Klassen Ruslan A, Valihrach Lukas, Romanyuk Nataliya
Department of Neuroregeneration, Institute of Experimental Medicine CAS, Prague, Czechia.
Department of Neuroscience, 2nd Medical Faculty, Charles University, Prague, Czechia.
Front Neurosci. 2025 Jun 26;19:1601101. doi: 10.3389/fnins.2025.1601101. eCollection 2025.
Oxidative stress (OS) is a key contributor to secondary damage following spinal cord injury (SCI), leading to neural stem cell (NSC) dysfunction and apoptosis. MicroRNA-20a (miR-20a) is upregulated after SCI and plays a role in regulating apoptosis and survival pathways. This study explores the therapeutic potential of miR-20a inhibition in mitigating OS-induced damage in NSCs.
Human iPSC-derived NSCs were subjected to oxidative stress by exposure to 100 µM hydrogen peroxide (HO) for 2 hours, followed by treatment with a miR-20a inhibitor (100 nM) to attenuate the adverse effects. Metabolic activity was evaluated using the Alamar Blue assay. Apoptotic responses and miR-20a expression levels were assessed via flow cytometry, RT-qPCR, and Western blot analysis.
NSCs exposed to OS showed a marked reduction in metabolic activity. However, treatment with a miR-20a inhibitor over 72 h significantly improved cell survival and metabolic activity in a time-dependent manner compared to untreated stressed cells.
Our findings suggest that miR-20a inhibition mitigates OS-induced cytotoxicity and promotes NSC viability, presenting a potential therapeutic approach for enhancing neural tissue regeneration.
氧化应激（OS）是脊髓损伤（SCI）后继发性损伤的关键因素，导致神经干细胞（NSC）功能障碍和凋亡。微小RNA-20a（miR-20a）在SCI后上调，并在调节凋亡和生存途径中发挥作用。本研究探讨抑制miR-20a在减轻OS诱导的NSC损伤中的治疗潜力。
将人诱导多能干细胞衍生的NSC暴露于100μM过氧化氢（HO）中2小时以施加氧化应激，随后用miR-20a抑制剂（100 nM）处理以减轻不良反应。使用Alamar Blue测定法评估代谢活性。通过流式细胞术、RT-qPCR和蛋白质印迹分析评估凋亡反应和miR-20a表达水平。
暴露于OS的NSC代谢活性显著降低。然而，与未处理的应激细胞相比，用miR-20a抑制剂处理72小时以上可显著改善细胞存活和代谢活性，且呈时间依赖性。
我们的研究结果表明，抑制miR-20a可减轻OS诱导的细胞毒性并促进NSC活力，为增强神经组织再生提供了一种潜在的治疗方法。
