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| 人脐带间充质干细胞对Aβ1-42诱导SH-SY5Y细胞损伤的改善作用及机制研究 |
| Ameliorative effect and mechanism of human umbilical cord mesenchymal stem cells on Aβ-induced SH-SY5Y cell injury |
| 投稿时间：2024-07-01 修订日期：2024-07-01 |
| DOI： |
| 中文关键词: 间充质干细胞，阿尔茨海默病，细胞凋亡，细胞增殖，Apelin信号通路 |
| 英文关键词: Mesenchymal stem cells, Alzheimer’s disease, Apoptosis, Cell proliferation, Apelin signaling pathway |
| 基金项目:河南省医学科技攻关项目(242102310112,242102310144),河南省医学科技攻关计划联合共建项目(LHGJ20220858) |
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| 中文摘要: |
| 目的: 基于体外细胞实验探索人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)对Aβ1-42诱导SH-SY5Y细胞损伤的改善作用及相关机制。方法：提取hUC-MSCs并使用流式细胞术检测细胞表面标志物；油红O染色,茜素红S染色和阿利新蓝染色检测多向分化潜能。以SH-SY5Y细胞为对照组；使用10μM/ml的Aβ1-42处理SH-SY5Y细胞24 h设置为Aβ组；Aβ组SH-SY5Y细胞与hUC-MSCs等比例共培养24 h设置为MSC组。使用AnnexinⅤ-PI双染色法检测三组细胞凋亡,CCK8法检测三组细胞增殖。提取Aβ组和MSC组的总RNA行转录组测序,对结果数据进行差异表达分析和富集分析。实时定量PCR检测Aβ组和MSC组细胞中APLN和APLNR的mRNA水平表达。结果: 所提取的hUC-MSCs细胞表面阳性表达CD73、CD90和CD105,阴性表达CD11b、CD19、CD34、CD45和HLA-DR,且具有良好的成骨分化,成脂分化和成软骨分化潜能。与对照组相比,Aβ组SH-SY5Y细胞数量减少、形态皱缩,早期凋亡和晚期凋亡增加,存活率下降。与Aβ组相比,MSC组细胞形态得到恢复,早期凋亡和晚期凋亡减少,存活率增加。差异表达分析到287个上调的差异表达基因和142个下调的差异表达基因,上调差异表达基因可能参与神经活性配体-受体相互作用、Apelin信号通路和PI3K?Akt信号通路等信号通路。实时定量PCR结果显示MSC组细胞中APLN和APLNR的相对表达水平较Aβ组显著升高(P<0.05)。结论：本研究发现hUC-MSCs显著改善了Aβ1-42诱导的SH-SY5Y细胞损伤,其作用机制可能是通过调控Apelin信号通路发挥的。 |
| 英文摘要: |
| Objective: This study aims to investigate the protective effects and underlying mechanisms of human umbilical cord mesenchymal stem cells (hUC-MSCs) on Aβ1-42-induced damage in SH-SY5Y cells through in vitro experiments. Methods: hUC-MSCs were isolated and cell surface markers were detected using flow cytometry. Multidirectional differentiation potential of hUC-MSCs was detected using Oil Red O staining, Alizarin Red S staining and Alizarin Blue staining. SH-SY5Y cells were used 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 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 detected by real-time quantitative PCR. Results: The cell surface of the extracted hUC-MSCs positively expressed CD73, CD90 and CD105, and negatively expressed CD11b, CD19, CD34, CD45 and HLA-DR. The extracted hUC-MSCs cells have good osteogenic differentiation, lipogenic differentiation and chondrogenic differentiation potentials. Compared with the control group, SH-SY5Y cells in the Aβ group were reduced in number, 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 PI3K-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: In this study, we found that hUC-MSCs significantly ameliorated Aβ1-42-induced SH-SY5Y cell injury, and the mechanism of action may be exerted through the regulation of the Apelin signaling pathway. |
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