Mudyanselage Anusha W, Wijamunige Buddhika C, Kocoń Artur, Turner Ricky, McLean Denise, Morentin Benito, Callado Luis F, Carter Wayne G
Clinical Toxicology Research Group, School of Medicine, University of Nottingham, Royal Derby Hospital Centre, Uttoxeter Road, Derby DE22 3DT, UK.
Department of Export Agriculture, Faculty of Agricultural Sciences, Sabaragamuwa University of Sri Lanka, Belihuloya 70140, Sri Lanka.
Antioxidants (Basel). 2024 May 8;13(5):580. doi: 10.3390/antiox13050580.
Alcohol is toxic to neurons and can trigger alcohol-related brain damage, neuronal loss, and cognitive decline. Neuronal cells may be vulnerable to alcohol toxicity and damage from oxidative stress after differentiation. To consider this further, the toxicity of alcohol to undifferentiated SH-SY5Y cells was compared with that of cells that had been acutely differentiated. Cells were exposed to alcohol over a concentration range of 0-200 mM for up to 24 h and alcohol effects on cell viability were evaluated via MTT and LDH assays. Effects on mitochondrial morphology were examined via transmission electron microscopy, and mitochondrial functionality was examined using measurements of ATP and the production of reactive oxygen species (ROS). Alcohol reduced cell viability and depleted ATP levels in a concentration- and exposure duration-dependent manner, with undifferentiated cells more vulnerable to toxicity. Alcohol exposure resulted in neurite retraction, altered mitochondrial morphology, and increased the levels of ROS in proportion to alcohol concentration; these peaked after 3 and 6 h exposures and were significantly higher in differentiated cells. Protein carbonyl content (PCC) lagged behind ROS production and peaked after 12 and 24 h, increasing in proportion to alcohol concentration, with higher levels in differentiated cells. Carbonylated proteins were characterised by their denatured molecular weights and overlapped with those from adult post-mortem brain tissue, with levels of PCC higher in alcoholic subjects than matched controls. Hence, alcohol can potentially trigger cell and tissue damage from oxidative stress and the accumulation of oxidatively damaged proteins.
酒精对神经元有毒性，可引发与酒精相关的脑损伤、神经元丢失和认知衰退。神经元细胞在分化后可能易受酒精毒性和氧化应激损伤。为进一步研究此问题，将酒精对未分化的SH-SY5Y细胞的毒性与急性分化后的细胞毒性进行了比较。细胞暴露于浓度范围为0 - 200 mM的酒精中长达24小时，并通过MTT和LDH测定评估酒精对细胞活力的影响。通过透射电子显微镜检查对线粒体形态的影响，并使用ATP测量和活性氧（ROS）产生来检查线粒体功能。酒精以浓度和暴露持续时间依赖性方式降低细胞活力并消耗ATP水平，未分化细胞对毒性更敏感。酒精暴露导致神经突回缩、线粒体形态改变，并使ROS水平与酒精浓度成比例增加；这些在暴露3小时和6小时后达到峰值，且在分化细胞中显著更高。蛋白质羰基含量（PCC）滞后于ROS产生，并在12小时和24小时后达到峰值，与酒精浓度成比例增加，在分化细胞中水平更高。羰基化蛋白质通过其变性分子量进行表征，并与成人死后脑组织中的蛋白质重叠，酒精中毒受试者的PCC水平高于匹配的对照组。因此，酒精可能会引发氧化应激导致的细胞和组织损伤以及氧化损伤蛋白质的积累。
