Studies On Myelination Of Oligodendrocyte-like And Its Conditioned Medium For Oxidative Stress Injury Repair

Background Neural stem cells(NSCs),as a kind of pluripotent stem cells,have the potential of self-renewal and multi-differentiation.Oligodendroglia cells are regulatory cells of the central nervous system,whose main physiological functions are to wrap neuronal axons to form myelin sheath and regulate the nervous system microenvironment through paracrine action to provide nutritional support,protect neurons and inhibit neuronal apoptosis.Excessive oxidative stress leads to neuronal damage,exacerbates the occurrence and development of neurodegenerative diseases,and often causes neurological dysfunction such as memory and cognition in patients.The source of primary OCs in clinical practice is difficult,nsc-derived OCs and the related active substances secreted by them can provide a continuous stream of biological materials for tissue engineering for the treatment of neurodegenerative diseases.The purpose of this study was to explore the differentiation of NSCs into Oligodendrocyte-like and verify their myelinating ability,and to study the repair effect of active substances secreted by OLs on neuronal damage,providing new ideas for the treatment of nerve loss diseases.Objectives NSCs were differentiated into OLs and neurons,and OLs and neurons were co-cultured to verify the myelinating ability of OLs,and the protective effect of OCM on H2O2-induced oxidative stress injury of nerve cells was further explored in vitro.Methods(1)NSCs differentiated into OLs and neurons and were identified by morphological and immunofluorescence tests.(2)OLs coculture with neurons and myelination was verified by confocal imaging in vitro.(3)Establishing the SH-SY5 Y cell oxidative damage model induced by H2O2.(4)The protective effect of OCM on SH-SY5 Y cells was observed by flow cytometry,fluorescence microscopy and microplate reader.(5)Western blot,immunofluorescence and Sirt1 inhibitor EX527 were used to explore the specific mechanism of OCM improving oxidative stress injury of SH-SY5 Y cells.Results(1)OLs from human NSCs showed typical oligodendrocyte morphology,with high expression of oligodendrocyte markers Olig2 and Sox 10.The morphology of NSC-Neu was typical,and Tuj-1 immunofluorescence staining was positive for neuron marker.(2)OLs were observed by confocal microscopy to wrap neuronal axons and form myelin-like structures.(3)Compared with H2O2 group,OCM treatment significantly restored the morphology and vitality of SH-SY5 Y cells(P < 0.001)and reduced the release of LDH(P < 0.001).(4)Compared with H2O2 group,OCM treatment reduced ROS toxicity(P < 0.001),increased SOD(P < 0.01)and GSH levels(P < 0.001),decreased MDA content(P < 0.001),and increased mitochondrial membrane potential.(5)Compared with H2O2 group,OCM alleviated the apoptosis of SH-SY5 Y cells induced by H2O2(P < 0.001),increased the expression of anti-apoptotic protein Bcl-2(P < 0.05)and decreased the expression of pro-apoptotic protein Bax(P < 0.05).(6)OCM treatment inhibited the decrease of Sirt1(P < 0.05)and PGC-1α(P < 0.001)protein expression in SH-SY5 Y cells induced by H2O2.(7)The protective effect of OCM on SH-SY5 Y cells was inhibited by Sirt1 specific inhibitor EX527.Conclusions(1)NSCs can differentiate into OLs and neurons through targeted culture,and OLs have the ability to myelinate in vitro.(2)OCM can improve H2O2-induced oxidative stress of SH-SY5 Y cells and reduce SH-SY5 Y cell apoptosis through Sirt1/PGC-1α pathway.