Sánchez-Sánchez Laura, Fernández Roberto, Ganfornina Maria Dolores, Astigarraga Egoitz, Barreda-Gómez Gabriel
Research and Development Department, IMG Pharma Biotech S.L, 48160 Derio, Spain.
Instituto de Biología y Genética Molecular (IBGM), Unidad de Excelencia, Universidad de Valladolid-CSIC, 47003 Valladolid, Spain.
Antioxidants (Basel). 2022 Dec 10;11(12):2440. doi: 10.3390/antiox11122440.
Cellular senescence is one of the main contributors to some neurodegenerative disorders. The early detection of senescent cells or their related effects is a key aspect in treating disease progression. In this functional deterioration, oxidative stress and lipid peroxidation play an important role. Endogenous antioxidant compounds, such as α-tocopherol (vitamin E), can mitigate these undesirable effects, particularly lipid peroxidation, by blocking the reaction between free radicals and unsaturated fatty acid. While the antioxidant actions of α-tocopherol have been studied in various systems, monitoring the specific effects on cell membrane lipids at scales compatible with large screenings has not yet been accomplished. Understanding the changes responsible for this protection against one of the consequences of senescence is therefore necessary. Thus, the goal of this study was to determinate the changes in the lipid environment of a Paraquat-treated human astrocytic cell line, as a cellular oxidative stress model, and the specific actions of the antioxidant, α-tocopherol, using cell membrane microarray technology, MALDI-MS and lipidomic analysis. The stress induced by Paraquat exposure significantly decreased cell viability and triggered membrane lipid changes, such as an increase in certain species of ceramides that are lipid mediators of apoptotic pathways. The pre-treatment of cells with α-tocopherol mitigated these effects, enhancing cell viability and modulating the lipid profile in Paraquat-treated astrocytes. These results demonstrate the lipid modulation effects of α-tocopherol against Paraquat-promoted oxidative stress and validate a novel analytical high-throughput method combining cell cultures, microarray technology, MALDI-MS and multivariate analysis to study antioxidant compounds against cellular senescence.
细胞衰老在某些神经退行性疾病中是主要促成因素之一。衰老细胞或其相关效应的早期检测是治疗疾病进展的关键方面。在这种功能衰退中,氧化应激和脂质过氧化起着重要作用。内源性抗氧化化合物,如α-生育酚(维生素E),可通过阻断自由基与不饱和脂肪酸之间的反应来减轻这些不良影响,尤其是脂质过氧化。虽然α-生育酚的抗氧化作用已在各种系统中进行了研究,但尚未在与大规模筛查兼容的尺度上监测其对细胞膜脂质的具体影响。因此,有必要了解这种针对衰老后果之一的保护作用所涉及的变化。因此,本研究的目的是使用细胞膜微阵列技术、基质辅助激光解吸电离质谱(MALDI-MS)和脂质组学分析,确定作为细胞氧化应激模型的百草枯处理的人星形胶质细胞系脂质环境的变化以及抗氧化剂α-生育酚的具体作用。百草枯暴露诱导的应激显著降低了细胞活力并引发了膜脂质变化,例如某些神经酰胺种类增加,而神经酰胺是凋亡途径的脂质介质。用α-生育酚对细胞进行预处理可减轻这些影响,提高细胞活力并调节百草枯处理的星形胶质细胞中的脂质谱。这些结果证明了α-生育酚对百草枯促进的氧化应激的脂质调节作用,并验证了一种结合细胞培养、微阵列技术、MALDI-MS和多变量分析来研究抗氧化化合物对抗细胞衰老的新型高通量分析方法。