Chiang Ming-Chang, Nicol Christopher J B, Yang Yu-Ping, Chiang Tairui, Yen Chiahui
Department of Life Science, College of Science and Engineering, Fu Jen Catholic University, New Taipei City 242, Taiwan.
Departments of Pathology & Molecular Medicine and Biomedical & Molecular Sciences, and Cancer Biology and Genetics Division, Sinclair Cancer Research Institute, Queen's University, Kingston, ON K7L 3N6, Canada.
Neuroscience. 2025 Mar 17;569:67-84. doi: 10.1016/j.neuroscience.2025.02.002. Epub 2025 Feb 3.
Exposure to PM is associated with neurotoxicity and mitochondrial dysfunction. Resveratrol, a natural polyphenol, has demonstrated antioxidant and neuroprotective properties. Still, its efficacy in mitigating PM-induced damage in human neural stem cells (hNSCs) and within a 3D scaffold system remains underexplored.
This study investigated the protective effects of resveratrol against PM-induced damage in hNSCs and within a 3D scaffold system.
Assess cell viability using MTT and LIVE/DEAD assays and measure caspase activity by fluorescence analysis. Quantify gene and protein expression of key regulatory pathways using qPCR and Western blotting. Then, mitochondrial function was analyzed by measuring ATP production, mitochondrial mass, maximal respiratory rate, COX activity, membrane potential, TEM, and immunofluorescence staining. In addition, 3D scaffolds created by the CELLINK INKREDIBLE bioprinter were used to study the effect of resveratrol on PM-induced hNSCs damage.
Resveratrol significantly improved cell viability and reduced caspase-3 and caspase-9 activities in PM-treated hNSCs. Resveratrol treatment upregulated TrKBR, PI3K, AKT, CREB, PPARα, PPARγ, SIRT1 and AMPK expression. It restored mitochondrial function by increasing ATP production, mitochondrial mass, maximal respiratory rate, COX activity, and membrane potential. Using a 3D scaffold demonstrated resveratrol's potential to maintain mitochondrial function and cellular health under PM exposure.
Resveratrol can effectively reduce neurotoxicity and mitochondrial dysfunction caused by PM in hNSCs. Its protective effects against PM-induced toxicity in hNSCs within a 3D bioprinted model highlight this study's translational potential. These findings emphasize its potential as a therapeutic agent against environmental neurotoxins and the development of neuroprotective strategies.
接触细颗粒物（PM）与神经毒性和线粒体功能障碍有关。白藜芦醇是一种天然多酚，已显示出抗氧化和神经保护特性。然而，其在减轻PM对人神经干细胞（hNSCs）的损伤以及在三维支架系统中的功效仍未得到充分研究。
本研究调查了白藜芦醇对PM诱导的hNSCs损伤以及在三维支架系统中的保护作用。
使用MTT和活/死细胞检测法评估细胞活力，并通过荧光分析测量半胱天冬酶活性。使用qPCR和蛋白质印迹法对关键调控通路的基因和蛋白质表达进行定量。然后，通过测量ATP生成、线粒体质量、最大呼吸速率、COX活性、膜电位、透射电子显微镜（TEM）和免疫荧光染色来分析线粒体功能。此外，使用CELLINK INKREDIBLE生物打印机创建的三维支架来研究白藜芦醇对PM诱导的hNSCs损伤的影响。
白藜芦醇显著提高了PM处理的hNSCs的细胞活力，并降低了caspase-3和caspase-9的活性。白藜芦醇处理上调了TrKBR、PI3K、AKT、CREB、PPARα、PPARγ、SIRT1和AMPK的表达。它通过增加ATP生成、线粒体质量、最大呼吸速率、COX活性和膜电位来恢复线粒体功能。使用三维支架证明了白藜芦醇在PM暴露下维持线粒体功能和细胞健康的潜力。
白藜芦醇可以有效降低PM在hNSCs中引起的神经毒性和线粒体功能障碍。其在三维生物打印模型中对PM诱导的hNSCs毒性的保护作用突出了本研究的转化潜力。这些发现强调了其作为抗环境神经毒素治疗剂和神经保护策略开发的潜力。
