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  

Oligodendrocytes-like conditional medium can improve oxidative stress damage of SH-SY5Y cells by regulating Sirt1/PGC-1α pathway

作　　者：牛冬冬 凌亚亭 吕孝瑞 王子瑜 胡嘉波[1] NIU Dongdong;LING Yating;LYU Xiaorui;WANG Ziyu;HU Jiabo(School of Medicine,Jiangsu University,Zhenjiang Jiangsu 212013;Health Clinical Laboratories,Health BioMed Co.Ltd.,Ningbo Zhejiang 315042,China)

机构地区：[1]江苏大学医学院,江苏镇江212013 [2]海尔施生物医药股份有限公司医学检验所,浙江宁波315042

出　　处：《江苏大学学报（医学版）》2023年第5期412-419,共8页Journal of Jiangsu University：Medicine Edition

基　　金：校企协同创新项目(20200605)。

摘　　要：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H 2O 2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H 2O 2诱导的SH-SY5Y细胞氧化应激,减少SH-SY5Y细胞凋亡。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 H 2O 2.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-1αprotein(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 H 2O 2-induced oxidative stress of SH-SY5Y cells and reduce SH-SY5Y cell apoptosis through Sirt1/PGC-1αpathway.

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-1α通路 

分 类 号：R741.05[医药卫生—神经病学与精神病学]