Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Pathumthani, 12120, Thailand.
Center of Excellence in Stem Research and Innovation, Thammasat University, Pathumthani, 12120, Thailand.
Sci Rep. 2023 Dec 27;13(1):22974. doi: 10.1038/s41598-023-49751-5.
Placenta-derived mesenchymal stem cells (PL-MSCs) have therapeutic potential in various clinical contexts due to their regenerative and immunomodulatory properties. However, with increasing age or extensive in vitro culture, their viability and function are gradually lost, thus restricting their therapeutic application. The primary cause of this deterioration is oxidative injury from free radicals. Therefore, enhancing cell viability and restoring cellular repair mechanisms of PL-MSCs in an oxidative stress environment are crucial in this context. Fucoxanthin, a carotenoid derived from brown seaweed, demonstrates antioxidant activity by increasing the production of antioxidant enzymes and lowering the levels of reactive oxygen species (ROS). This study aimed to determine whether fucoxanthin protects PL-MSCs from hydrogen peroxide (HO)-induced oxidative stress. After characterization, PL-MSCs were co-treated with fucoxanthin and HO for 24 h (co-treatment) or pre-treated with fucoxanthin for 24 h followed by HO for 24 h (pre-treatment). The effects of fucoxanthin on cell viability and proliferation were examined using an MTT assay. The expression of antioxidant enzymes, PI3K/Akt/Nrf-2 and intracellular ROS production were investigated in fucoxanthin-treated PL-MSCs compared to the untreated group. The gene expression and involvement of specific pathways in the cytoprotective effect of fucoxanthin were investigated by high-throughput NanoString nCounter analysis. The results demonstrated that co-treatment and pre-treatment with fucoxanthin restored the viability and proliferative capacity of PL-MSCs. Fucoxanthin treatment increased the expression of antioxidant enzymes in PL-MSCs cultured under oxidative stress conditions and decreased intracellular ROS accumulation. Markedly, fucoxanthin treatment could restore PI3K/Akt/Nrf-2 expression in HO-treated PL-MSCs. High-throughput analysis revealed up-regulation of genes involved in cell survival pathways, including cell cycle and proliferation, DNA damage repair pathways, and down-regulation of genes in apoptosis and autophagy pathways. This study demonstrated that fucoxanthin protects and rescues PL-MSCs from oxidative stress damage through the PI3K/Akt/Nrf-2 pathway. Our data provide the supporting evidence for the use of fucoxanthin as an antioxidant cytoprotective agent to improve the viability and proliferation capacity of PL-MSCs both in vitro and in vivo required to increase the effectiveness of MSC expansion for therapeutic applications.
胎盘间充质干细胞（PL-MSCs）具有再生和免疫调节特性，因此在各种临床环境中具有治疗潜力。然而，随着年龄的增长或体外广泛培养，其活力和功能逐渐丧失，从而限制了其治疗应用。这种恶化的主要原因是自由基引起的氧化损伤。因此，在这种情况下，增强 PL-MSCs 在氧化应激环境中的细胞活力并恢复细胞修复机制至关重要。褐藻来源的叶黄素是一种类胡萝卜素，通过增加抗氧化酶的产生和降低活性氧物种（ROS）的水平来发挥抗氧化活性。本研究旨在确定叶黄素是否能保护 PL-MSCs 免受过氧化氢（HO）诱导的氧化应激。在表征后，将 PL-MSCs 与叶黄素和 HO 共同处理 24 小时（共同处理）或先用叶黄素预处理 24 小时，然后再用 HO 处理 24 小时（预处理）。使用 MTT 测定法检查叶黄素对细胞活力和增殖的影响。与未处理组相比，研究了叶黄素处理的 PL-MSCs 中抗氧化酶、PI3K/Akt/Nrf-2 的表达和细胞内 ROS 产生。通过高通量 NanoString nCounter 分析研究了叶黄素的细胞保护作用中的基因表达和特定途径的参与。结果表明，共同处理和预处理叶黄素恢复了 PL-MSCs 的活力和增殖能力。叶黄素处理可增加氧化应激条件下培养的 PL-MSCs 中抗氧化酶的表达并减少细胞内 ROS 的积累。值得注意的是，叶黄素处理可恢复 HO 处理的 PL-MSCs 中 PI3K/Akt/Nrf-2 的表达。高通量分析显示，参与细胞存活途径的基因上调，包括细胞周期和增殖、DNA 损伤修复途径，以及凋亡和自噬途径的基因下调。本研究表明，叶黄素通过 PI3K/Akt/Nrf-2 途径保护和挽救 PL-MSCs 免受氧化应激损伤。我们的数据为使用叶黄素作为抗氧化细胞保护剂提供了支持证据，以提高 MSC 扩增的有效性，从而提高其在体外和体内治疗应用中的活力和增殖能力。
