Endothelial progenitor cells-secreted extracellular vesicles containing microRNA-93-5p confer protection against sepsis-induced acute kidney injury via the KDM6B/H3K27me3/TNF-α axis
Endothelial progenitor cells-secreted extracellular vesicles containing microRNA-93-5p confer protection against sepsis-induced acute kidney injury via the KDM6B/H3K27me3/TNF-α axis
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DOI：
10.1016/j.yexcr.2020.112173
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发表时间：
2020-10-15
影响因子：
3.7
通讯作者：
Yue, Lingju
中科院分区：
文献类型：
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作者：
He, Zhonghua;Wang, Haixia;Yue, Lingju
越来越多的证据表明microRNA（miRs）在脓毒症诱导的急性肾损伤（AKI）中的关键致病作用。靶细胞的功能通过将细胞包封的细胞外囊泡（Evs）释放到细胞外空间中来调节。本研究旨在阐明内皮祖细胞（EPC）衍生的Evs中含有miR-93 - 5p在脓毒症诱导的AKI中的临床意义以及生物学功能。我们首先通过脂多糖（LPS）处理建立细胞脓毒症诱导的AKI小鼠模型，并在模型中测试异位表达和消除实验。分离来源于miR-93 - 5p转染的EPCs的Evs（Evs/miR-93 - 5p抑制剂），并与HK 2细胞共培养，以探讨过表达miR-93 - 5p的EPC来源的Evs对LPS诱导的HK 2细胞损伤的影响。采用双荧光素酶报告基因检测miR-93 - 5p与赖氨酸（K）特异性脱甲基酶6B（KDM6B）的相互作用，并采用ChIP验证KDM6B与肿瘤坏死因子-α（TNF-α）的关系。采用盲肠结扎穿孔法（CLP）制备小鼠脓毒症模型，注射Ev/miR-93 - 5p抑制剂，观察其在体内的作用。结果发现，在LPS处理的HK2细胞中，miR-93 - 5p和组蛋白H3 Lys27三甲基化（H3K27me3）下调，而KDM6B上调。Ev/miR-93 - 5p抑制剂可增强LPS对HK2细胞的损伤作用。发现miR-93 - 5p直接靶向KDM6B。沉默KDM6B诱导H3K27me3，抑制TNF-α的活化，从而减弱LPS诱导的HK 2细胞损伤。EPC衍生的含有miR-93 - 5p的Evs减轻脓毒症小鼠中的多器官损伤、血管渗漏、炎症和细胞凋亡。结论：携带miR-93 - 5p的EPC源性Ev对脓毒症大鼠AKI的保护作用可能是通过调节KDM6BH/3K27me3/TNF-α轴介导的。
The pivotal pathogenetic role of microRNAs (miRs) in sepsis-induced acute kidney injury (AKI) has been demonstrated in mounting evidence. The functions of the target cells are regulated through the release of cells-encapsulated extracellular vesicles (Evs) into the extracellular space. The present study aims to elucidate the clinical significance as well as biological function of the endothelial progenitor cell (EPC)-derived Evs containing miR-93-5p in sepsis-induced AKI. We first established a cellular sepsis-induced AKI mouse model by treatment with lipopolysaccharide (LPS), and tested ectopic expression and depletion experiments in the model. Evs derived from miR-93-5p inhibitor-transfected EPCs (Evs/miR-93-5p inhibitor) were isolated, and co-cultured with HK2 cells to explore the effects of EPC-derived Evs overexpressing miR-93-5p on LPS-induced HK2 cell injury. The interaction between miR-93-5p and lysine (K)-specific demethylase 6B (KDM6B) was identified using dual-luciferase reporter assay, and ChIP was used to validate the relationship between KDM6B and tumor necrosis factor-alpha (TNF-alpha). Mice were made septic by cecal ligation and puncture (CLP), and then injected with Ev/ miR-93-5p inhibitor to explore its functions in vivo. The results found that miR-93-5p and histone H3 Lys27 trimethylation (H3K27me3) were downregulated while KDM6B was upregulated in LPS-treated HK2 cells. EPCderived Evs alleviated LPS-induced HK2 cell injury, while Ev/miR-93-5p inhibitor potentiated the cell injury in vitro. miR-93-5p was found to directly target KDM6B. Silencing KDM6B induced H3K27me3, inhibiting the activation of TNF-alpha, thereby weakening LPS-induced HK2 cell injury. EPC-derived Evs containing miR-93-5p attenuated multiple organ injury, vascular leakage, inflammation, and apoptosis in septic mice. In conclusion, the present study demonstrated that endothelial protection from EPC-derived Evs carrying miR-93-5p in sepsisinduced AKI, which was mediated by regulation KDM6BH/3K27me3/TNF-alpha axis.