脐带间充质干细胞条件培养基的抗氧化活性和抗铁死亡作用

Anti-oxidative activities and anti-ferroptosis of conditioned medium from umbilical cord mesenchymal stem cells.

作者信息

Liu Yu-Bin, Zhang Lin

机构信息

Division of (Bio) Pharmaceutics, Institute of Zhejiang University - Quzhou, Zhejiang, China.

The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Zhejiang, China.

出版信息

In Vitro Cell Dev Biol Anim. 2023 Oct;59(9):658-664. doi: 10.1007/s11626-023-00816-9. Epub 2023 Nov 3.

Abstract

The therapeutic effect of MSC is closely related to its antioxidant capacity. There is no uniform standard for evaluating the antioxidant capacity of MSC. In this study, we compared the antioxidant capacity of control medium (CON) and conditioned medium (CM) from umbilical cord mesenchymal stem cells cultured for 48 h, about total antioxidant capacity, DPPH scavenging capacity, O2- and hydroxyl radical inhibiting capacity, and the detection of antioxidant enzymes including superoxide dismutase, glutathione peroxidase, and catalase, and resistance to cellular oxidative damage caused by H2O2, SNAP, erastin, and RSL3. The results showed that CM had better DPPH scavenging capacity than CON. No significant differences were observed in antioxidant enzymes. CM did not resist the oxidative damage induced by H2O2 and SNAP, but it had a strong resistance to ferroptosis induced by erastin and RSL3, indicating that CM had excellent resistance to cell lipid peroxidation. CM could improve the cell shrinkage morphology induced by ferroptosis and reduce the production of lipid ROS. qPCR experiments proved that CM improved and regulated multiple pathways of ferroptosis, including genes related to iron metabolism such as FPN, FTH1, TFRC, and IREB2, and redox regulatory genes such as GPX4, AIFM2, DHODH, and TP53, and increased the antioxidant-related transcription factors NRF2 and ATF4.

摘要

间充质干细胞(MSC)的治疗效果与其抗氧化能力密切相关。目前尚无评估MSC抗氧化能力的统一标准。在本研究中,我们比较了培养48小时的脐带间充质干细胞的对照培养基(CON)和条件培养基(CM)的抗氧化能力,包括总抗氧化能力、DPPH清除能力、超氧阴离子和羟自由基抑制能力,以及对抗氧化酶(包括超氧化物歧化酶、谷胱甘肽过氧化物酶和过氧化氢酶)的检测,以及对由过氧化氢、硝普钠、艾拉司亭和RSL3引起的细胞氧化损伤的抗性。结果表明,CM的DPPH清除能力优于CON。抗氧化酶方面未观察到显著差异。CM不能抵抗过氧化氢和硝普钠诱导的氧化损伤,但对艾拉司亭和RSL3诱导的铁死亡具有较强的抗性,表明CM对细胞脂质过氧化具有优异的抗性。CM可以改善铁死亡诱导的细胞收缩形态,并减少脂质活性氧的产生。定量聚合酶链反应(qPCR)实验证明,CM改善并调节了铁死亡的多个途径,包括与铁代谢相关的基因,如铁转运蛋白1(FPN)、铁蛋白重链1(FTH1)、转铁蛋白受体1(TFRC)和铁反应元件结合蛋白2(IREB2),以及氧化还原调节基因,如谷胱甘肽过氧化物酶4(GPX4)、凋亡诱导因子线粒体相关2(AIFM2)、二氢乳清酸脱氢酶(DHODH)和肿瘤蛋白p53(TP53),并增加了抗氧化相关转录因子核因子E2相关因子2(NRF2)和活化转录因子4(ATF4)。

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