来自远程缺血预处理血清的外泌体微小RNA-126通过下调DNMT3B参与SH-SY5Y细胞的缺氧耐受。
Exosomal MicroRNA-126 from RIPC Serum Is Involved in Hypoxia Tolerance in SH-SY5Y Cells by Downregulating DNMT3B.
作者信息
Cui Junhe, Liu Na, Chang Zhehan, Gao Yongsheng, Bao Mulan, Xie Yabin, Xu Wenqiang, Liu Xiaolei, Jiang Shuyuan, Liu You, Shi Rui, Xie Wei, Jia Xiaoe, Shi Jinghua, Ren Changhong, Gong Kerui, Zhang Chunyang, Bade Rengui, Shao Guo, Ji Xunming
机构信息
Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Inner Mongolia, PRC; Biomedicine Research Center, Basic Medical College and Baotou Medical College of the Neuroscience Institute, Baotou Medical College, Inner Mongolia, PRC; Beijing Key Laboratory of Hypoxic Conditioning Translational Medicine, Xuanwu Hospital, Capital Medical University, Beijing, PRC.
Inner Mongolia Key Laboratory of Hypoxic Translational Medicine, Baotou Medical College, Inner Mongolia, PRC; Biomedicine Research Center, Basic Medical College and Baotou Medical College of the Neuroscience Institute, Baotou Medical College, Inner Mongolia, PRC.
出版信息
Mol Ther Nucleic Acids. 2020 Jun 5;20:649-660. doi: 10.1016/j.omtn.2020.04.008. Epub 2020 Apr 25.
Ischemic tolerance in the brain can be induced by transient limb ischemia, and this phenomenon is termed remote ischemic preconditioning (RIPC). It still remains elusive how this transfer of tolerance occurs. Exosomes can cross the blood-brain barrier, and some molecules may transfer neuroprotective signals from the periphery to the brain. Serum miRNA-126 is associated with ischemic stroke, and exosomal miRNA-126 has shown protective effects against acute myocardial infarction. Therefore, this study aims to explore whether exosomal miRNA-126 from RIPC serum can play a similar neuroprotective role. Exosomes were isolated from the venous serum of four healthy young male subjects, both before and after RIPC. Exosomal miRNA-126 was measured by real-time PCR. The miRNA-126 target sequence was predicted by bioinformatics software. SH-SY5Y neuronal cells were incubated with exosomes, and the cell cycle was analyzed by flow cytometry. The expression and activity of DNA methyltransferase (DNMT) 3B, a potential target gene of miRNA-126, were examined in SH-SY5Y cells. The cell viability of SH-SY5Y cells exposed to oxygen-glucose deprivation (OGD) was also investigated. To confirm the association between miRNA-126 and DNMT3B, we overexpressed miRNA-126 in SH-SY5Y cells using lentiviral transfection. miRNA-126 expression was upregulated in RIPC exosomes, and bioinformatics prediction showed that miRNA-126 could bind with DNMT3B. DNMT levels and DNMT3B activity were downregulated in SH-SY5Y cells incubated with RIPC exosomes. After overexpression of miRNA-126 in SH-SY5Y cells, global methylation levels and DNMT3B gene expression were downregulated in these cells, consistent with the bioinformatics predictions. RIPC exosomes can affect the cell cycle and increase OGD tolerance in SH-SY5Y cells. RIPC seems to have neuroprotective effects by downregulating the expression of DNMTs in neural cells through the upregulation of serum exosomal miRNA-126.
短暂性肢体缺血可诱导大脑产生缺血耐受，这种现象被称为远程缺血预处理（RIPC）。耐受是如何发生转移的仍不清楚。外泌体能够穿过血脑屏障，一些分子可能将神经保护信号从外周传递至大脑。血清miRNA-126与缺血性中风有关，外泌体miRNA-126已显示出对急性心肌梗死的保护作用。因此，本研究旨在探讨RIPC血清中的外泌体miRNA-126是否能发挥类似的神经保护作用。在RIPC前后，从四名健康年轻男性受试者的静脉血清中分离出外泌体。通过实时PCR检测外泌体miRNA-126。利用生物信息学软件预测miRNA-126的靶序列。将SH-SY5Y神经细胞与外泌体共同孵育，通过流式细胞术分析细胞周期。检测miRNA-126的潜在靶基因DNA甲基转移酶（DNMT）3B在SH-SY5Y细胞中的表达及活性。还研究了暴露于氧-葡萄糖剥夺（OGD）的SH-SY5Y细胞的活力。为了证实miRNA-126与DNMT3B之间的关联，我们使用慢病毒转染在SH-SY5Y细胞中过表达miRNA-126。RIPC外泌体中miRNA-126表达上调，生物信息学预测显示miRNA-126可与DNMT3B结合。与RIPC外泌体共同孵育的SH-SY5Y细胞中，DNMT水平和DNMT3B活性下调。在SH-SY5Y细胞中过表达miRNA-126后，这些细胞中的整体甲基化水平和DNMT3B基因表达下调，这与生物信息学预测结果一致。RIPC外泌体可影响细胞周期并增加SH-SY5Y细胞对OGD的耐受性。RIPC似乎通过上调血清外泌体miRNA-126来下调神经细胞中DNMTs的表达，从而发挥神经保护作用。
Zotero 插件