少突胶质样细胞条件培养液通过调节Sirt1/PGC-1α通路改善SH-SY5Y细胞氧化应激损伤

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目的： 研究人神经干细胞来源的少突胶质样细胞条件培养液（oligodendrocytes-like conditioned medium，OCM）对人神经元氧化应激损伤的修复作用及可能机制。方法: 人神经干细胞分化为少突胶质样细胞（oligodendrocytes-like，OLs），免疫荧光等方法鉴定OLs，体外观察其髓鞘形成能力。构建H2O2诱导的人神经母细胞瘤细胞（SH-SY5Y）氧化损伤模型，流式细胞术测定活性氧水平及细胞凋亡，荧光探针检测线粒体膜电位，化学发光法检测还原型谷胱甘肽（GSH）、丙二醛（MDA）、超氧化物歧化酶（SOD）水平。蛋白质印迹法检测Sirt1/PGC-1α通路蛋白水平。结果: 人神经干细胞源少突胶质样细胞（human neural stem cell-derived oligodendrocytes-like，hNSC-OLs）呈现典型的少突胶质细胞形态，表达少突胶质细胞标志物Olig2和SOX10，在体外包裹轴突形成髓鞘。OCM处理显著提高了受损SH-SY5Y细胞活力（P＜0.01），活性氧水平明显降低（P＜0.01），线粒体膜电位升高，GSH、SOD水平升高（P＜0.01），MDA含量下降（P＜0.01），凋亡水平下降（P＜0.01）。OCM处理提高了Sirt1和PGC-1α蛋白水平（P＜0.05），并能被抑制剂EX527特异抑制（P＜0.05）。结论: hNSC-OLs体外具有成髓鞘能力，OCM可以通过Sirt1/PGC-1α通路改善H2O2诱导的SHSY5Y细胞氧化应激，减少SH-SY5Y细胞凋亡。

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	牛冬冬1
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	胡嘉波1

关键词 ：神经干细胞, 髓鞘形成, 少突胶质样细胞条件培养液, 氧化应激, Sirt1/PGC-1&alpha, 通路

Abstract：Objective： To explore the repair effect of oligodendrocyteslike conditioned medium (OCM) derived from human neural stem cells (hNSCs) on human neuron and its possible mechanism. Methods： Human neural stem cells differentiated into oligodendrocytes-like (OLs), which were identified by immunofluorescence and their myelination ability was observed in vitro. The oxidative damage model of SH-SY5Y cells was induced by H2O2. Reactive oxygen species (ROS) and cell apoptosis were determined by flow cytometry. Mitochondrial membrane potential was detected by fluorescence microscopy. SOD, GSH and MDA were detected by chemiluminescence. The protein level of Sirt1/PGC-1α was determined by Western blotting. Results： Human neural stem cellderived oligodendrocytes-like (hNSC-OLs) have a typical oligodendrocyte morphology and express oligodendrocyte markers Olig2 and SOX10, the formation of myelin sheath around axons was observed in vitro. OCM significantly increased the vitality of the damaged cells (P<0.01), decreased ROS level (P<0.01), increased mitochondrial membrane potential, increased GSH, SOD level (P<0.01), decreased MDA content (P<0.01), and decreased apoptosis level (P<0.01). OCM treatment increased the expression levels of Sirt1 and PGC-1α protein (P<0.05), and was specifically inhibited by the inhibitor EX527 (P<0.05). Conclusion： hNSC-OLs have myelination ability in vitro, and its conditioned medium can improve H2O2induced oxidative stress of SH-SY5Y cells and reduce SHSY5Y cell apoptosis through Sirt1/PGC-1α pathway.

收稿日期: 2023-02-18

基金资助:校企协同创新项目（20200605）

通讯作者: 胡嘉波(通讯作者)，教授，博士生导师，E-mail: hu@ujs.edu.cn

作者简介: 牛冬冬（1995—），男，硕士研究生；

引用本文:

牛冬冬1，凌亚亭1，吕孝瑞1，王子瑜2，胡嘉波1. 少突胶质样细胞条件培养液通过调节Sirt1/PGC-1α通路改善SH-SY5Y细胞氧化应激损伤[J]. 江苏大学学报:医学版, 2023, 33(05): 412-419. NIU Dongdong1, LING Yating1, LYU Xiaorui1, WANG Ziyu2, HU Jiabo1. Oligodendrocytes-like conditional medium can improve oxidative stress damage of SH-SY5Y cells by regulating Sirt1/PGC-α pathway#br#. Journal of Jiangsu University(Medicine Edition), 2023, 33(05): 412-419.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2023/V33/I05/412

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