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 Aβ1-42-induced SH-SY5Y Cell Injury

作　　者：唐吉祥 邵一鸣 靳冉冉 孔宁 马保东 张辉 TANG Ji-xiang;SHAO Yi-ming;JIN Ran-ran;KONG Ning;MA Bao-dong;ZHANG Hui(Xinxiang Medical University,Xinxiang,Henan,453000,China;Department of Neurosurgery,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan,450001,China;Center of Stem Cell and Regenerative Medicine,Zhengzhou Central Hospital Affiliated to Zhengzhou University,Zhengzhou,Henan,450001,China)

机构地区：[1]新乡医学院,河南新乡453000 [2]郑州大学附属郑州中心医院神经外科,河南郑州450001 [3]郑州大学附属郑州中心医院干细胞再生医学转化中心,河南郑州450001

出　　处：《现代生物医学进展》2024年第23期4421-4427,共7页Progress in Modern Biomedicine

基　　金：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)。

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: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。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 10μM/mL of Aβ1-42 for 24 h set to the Aβgroup.SH-SY5Y cells of Aβgroup 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 Aβand 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 Aβand MSC groups were detected by real-time quantitative PCR.Results:The extracted hUC-MSCs were positive for CD73,CD90 and CD105 but negative for CD14,CD19,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 Aβgroup reduced,wrinkled in morphology,increased in early apoptosis and late apoptosis,and decreased in survival rate.Compared with the Aβgroup,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

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 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R742[医药卫生—基础医学]