Chu Che-Sheng, Chen Ying-Tso, Sun Wei-Chih, Liang Wei-Zhe
Department of Psychiatry, Kaohsiung Veterans General Hospital, Kaohsiung, 813414, Taiwan.
Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, 807378, Taiwan.
Mol Biol Rep. 2025 Apr 15;52(1):391. doi: 10.1007/s11033-025-10481-8.
The importance of fatty acids in human health and their potential in treating various brain diseases is increasingly acknowledged. Research indicates that ultra-long-chain fatty acids adversely affect dietary habits, while omega (ω)-3 polyunsaturated fatty acids confer health benefits. Eicosapentaenoic acid (EPA), an ω-3 polyunsaturated fatty acid, manifests diverse protective activities, including anti-oxidative effects and the attenuation of brain diseases. Previous studies have suggested that EPA can alleviate oxidative stress and forestall diseases stemming from oxidative damage. Nevertheless, EPA's precise antioxidant mechanism and signaling pathway in human astrocytes remain elusive. To address this knowledge gap, we established an HO-induced oxidative damage model in Gibco Human Astrocytes (GHA cells) and elucidated the underlying mechanisms and signaling pathways.
Our assessments included cell viability through the CCK-8 assay, morphological examination via microscopy, ROS quantification using the DCFH-DA fluorescent probe, GSH content evaluation with the CMF-DA fluorescent probe, and protein expression analysis for antioxidant and apoptotic markers through Western blotting. The results showed that pretreatment with 3 µM of EPA countered the cytotoxicity, ROS production, and GSH depletion caused by HO (250 µM) in GHA cells. Additionally, EPA pretreatment effectively reduced the cytotoxicity and oxidative stress resulting from HO by modulating the Nrf2/HO-1/NQO1 and Bax/Bcl-2/caspase-9/caspase-3 signaling pathways in GHA cells.
These findings enhance our understanding of EPA's antioxidant mechanisms in the oxidative stress model of human astrocytes, illuminate the interplay between antioxidant and apoptotic signals, and offer promise for exploring potential preventive and therapeutic interventions for brain diseases.
脂肪酸对人类健康的重要性及其在治疗各种脑部疾病方面的潜力日益得到认可。研究表明，超长链脂肪酸会对饮食习惯产生不利影响，而ω-3多不饱和脂肪酸则有益健康。二十碳五烯酸（EPA）作为一种ω-3多不饱和脂肪酸，具有多种保护作用，包括抗氧化作用以及对脑部疾病的缓解作用。先前的研究表明，EPA可以减轻氧化应激并预防由氧化损伤引起的疾病。然而，EPA在人星形胶质细胞中的精确抗氧化机制和信号通路仍不清楚。为了填补这一知识空白，我们在Gibco人星形胶质细胞（GHA细胞）中建立了HO诱导的氧化损伤模型，并阐明了其潜在机制和信号通路。
我们的评估包括通过CCK-8法检测细胞活力、通过显微镜进行形态学检查、使用DCFH-DA荧光探针定量ROS、使用CMF-DA荧光探针评估GSH含量，以及通过蛋白质印迹法分析抗氧化和凋亡标志物的蛋白质表达。结果表明，用3µM的EPA预处理可对抗HO（250µM）在GHA细胞中引起的细胞毒性、ROS产生和GSH消耗。此外，EPA预处理通过调节GHA细胞中的Nrf2/HO-1/NQO1和Bax/Bcl-2/caspase-9/caspase-3信号通路，有效降低了HO引起的细胞毒性和氧化应激。
这些发现加深了我们对EPA在人星形胶质细胞氧化应激模型中的抗氧化机制的理解，阐明了抗氧化和凋亡信号之间的相互作用，并为探索脑部疾病的潜在预防和治疗干预措施提供了希望。
