Ameliorative Effect and Mechanism of Human Umbilical Cord Mesenchymal Stem Cells on Aβ1-42-induced SH-SY5Y Cell Injury
Objective:To investigate the protective effects and mechanisms of human umbilical cord mesenchymal stem cells(hUC-MSCs)on Aβ1-42-induced damage in SH-SY5Y cells by in vitro experiments.Methods:hUC-MSCs were isolated and cell surface markers were detected by using flow cytometry.Multidirectional differentiation potential of hUC-MSCs was detected by using Oil Red O staining,Alizarin Red S staining and Alizarin Blue staining.SH-SY5Y cells were chosen as the control group,SH-SY5Y cells were treated with 10 μM/mL of Aβ1-42 for 24 h set to the Aβ group.SH-SY5Y cells of Aβ group were co-cultured with hUC-MSCs in equal proportions for 24 h set as MSC group.Apoptosis was detected in three groups of cells by using Annexin V-PI double staining,and cell proliferation was detected in three groups of cells using the CCK8 assay.Total RNA from the Aβ and MSC groups was extracted for transcriptome sequencing,and the resulting data were analyzed for differential expression and enrichment.Expression of APLN and APLNR at mRNA level in cells of Aβ and MSC groups were detected by real-time quantitative PCR.Results:The extracted hUC-MSCs were positive for CD73,CD90 and CD 105 but negative for CD 14,CD 19,CD34,CD45 and HLA-DR.The hUC-MSCs have good potential for osteogenic differentiation,lipogenic differentiation and chondrogenic differentiation.Compared with that in the control group,SH-SY5Y cells in the Aβ group reduced,wrinkled in morphology,increased in early apoptosis and late apoptosis,and decreased in survival rate.Compared with the Aβ group,cell morphology was restored,early apoptosis and late apoptosis were reduced,and survival was increased in the MSC group.Differential expression was analyzed to 287 up-regulated and 142 down-regulated differentially expressed genes.Up-regulated differentially expressed genes may be involved in signaling pathways such as neuroactive ligand-receptor interactions,Apelin signaling pathway and P13K-Akt signaling pathway.The results of real-time quantitative PCR showed that the relative expression levels of APLN and APLNR in the cells of the MSC group were significantly higher than those of the Aβ group.Conclusions:The hUC-MSCs significantly ameliorated Aβ1-42-induced SH-SY5Y cell injury,and the mechanism may be exerted via the Apelin signaling pathway.
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