Zang Huifen, Wang Zhaohui, Wu Qingqing, Shi Lei, Chen Ge
Department of Obstetrics, The NO. 8 People's Hospital of Qingdao, Qingdao, 266100, China.
Cell Tissue Bank. 2023 Dec;24(4):769-778. doi: 10.1007/s10561-023-10095-z. Epub 2023 May 24.
Mesenchymal stem cells (MSCs) derived extracellular vesicles, which have been shown to possess therapeutic effects for many diseases. However, how hypoxic conditions would affect exosomal microRNA expression in human umbilical cord MSCs (hUC-MSCs) is currently not investigated. This study aims to investigate the potential function of in vitro microRNAs of hUC-MSC cultured under normoxic and hypoxic conditions. Extracellular vesicles secreted from hUC-MSCs cultured in normoxic (21% O) and hypoxic (5% O) conditions were collected for microRNA identification. Zeta View Laser Scattering and transmission electron microscopy were used to observe the size and morphology of extracellular vesicles. qRT-PCR was performed to measure the expression of related microRNAs. The Gene Ontology and KEGG pathway were used to predict the function of microRNAs. Finally, the effects of hypoxia on the expression of related mRNAs and cellular activity were examined. This study identified 35 upregulated and 8 downregulated microRNAs in the hypoxia group. We performed target genes analysis to explore the potential function of these microRNA upregulated in the hypoxia group. Significant enrichment of the cell proliferation, pluripotency of stem cells, MAPK, Wnt, and adherens junction pathways were observed in the GO and KEGG pathways. Under hypoxic conditions, the expression levels of 7 target genes were lower than that of the normal environment. In conclusion, this study demonstrated for the first time that microRNA expression in extracellular vesicles of human umbilical vein stem cells cultured under hypoxia is different from that under normal conditions, and these microRNAs may be markers for detecting hypoxia.
间充质干细胞（MSCs）衍生的细胞外囊泡已被证明对多种疾病具有治疗作用。然而，低氧条件如何影响人脐带间充质干细胞（hUC-MSCs）中外泌体微小RNA的表达目前尚未得到研究。本研究旨在探讨在常氧和低氧条件下培养的hUC-MSCs体外微小RNA的潜在功能。收集在常氧（21% O₂）和低氧（5% O₂）条件下培养的hUC-MSCs分泌的细胞外囊泡用于微小RNA鉴定。使用Zeta View激光散射和透射电子显微镜观察细胞外囊泡的大小和形态。进行qRT-PCR以测量相关微小RNA的表达。使用基因本体论（Gene Ontology）和京都基因与基因组百科全书（KEGG）通路来预测微小RNA的功能。最后，检测低氧对相关mRNA表达和细胞活性的影响。本研究在低氧组中鉴定出35个上调的微小RNA和8个下调的微小RNA。我们进行了靶基因分析以探索低氧组中上调的这些微小RNA的潜在功能。在基因本体论和KEGG通路中观察到细胞增殖、干细胞多能性、丝裂原活化蛋白激酶（MAPK）、Wnt和黏附连接通路的显著富集。在低氧条件下，7个靶基因的表达水平低于正常环境。总之，本研究首次证明，低氧条件下培养的人脐静脉干细胞细胞外囊泡中的微小RNA表达与正常条件下不同，这些微小RNA可能是检测低氧的标志物。
