miR-98 protects endothelial cells against hypoxia/reoxygenation induced-apoptosis by targeting caspase-3
miR-98 protects endothelial cells against hypoxia/reoxygenation induced-apoptosis by targeting caspase-3
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DOI：
10.1016/j.bbrc.2015.09.058
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发表时间：
2015-11-20
影响因子：
3.1
通讯作者：
Yu, Guang
中科院分区：
文献类型：
--
作者：
Li, He-wen;Meng, Yan;Yu, Guang
内皮功能障碍是肾缺血再灌注损伤的主要病理生理过程之一。我们之前的微阵列研究表明，缺血再灌注损伤（IRI）肾脏中 miR-98 上调。本研究旨在探讨 miR-98 是否参与缺氧和复氧 (H/R) 条件下内皮细胞凋亡的调节。测量小鼠 IRI 肾和 H/R HUVEC 中 miR-98 的动态变化。用 HIF-1 α siRNA 处理 HUVEC，以研究 HIF-1 α 对 miR-98 表达的作用。通过生物信息学分析预测miR-98的潜在靶基因。用miR-98模拟物或抑制剂转染HUVEC，以证实miR-98对靶基因表达和缺氧诱导的细胞凋亡的作用。最终通过双荧光素酶报告基因检测证实了目的基因。 IRI 和 H/R 均诱导缺血肾和缺氧 HUVEC 中 miR-98 的显着上调。 HIF-1 α siRNA 显着下调正常和缺氧 HUVEC 中 miR-98 的表达。 miR-98的假定靶基因包括IL-6、IL-10和caspase-3。 MiR-98 模拟物显着抑制 HUVEC 中 caspase-3 的表达，而抗 miR-98 显着上调其表达。但转染miRNA后未观察到IL-6和IL-10水平的变化。 miR-98 保护 HUVEC 免受缺氧诱导的细胞凋亡，而抗 miR-98 则具有相反的作用。此外，双荧光素酶报告基因测定证实，miR-98 通过靶向 caspase-3 的 3' 非翻译区来降低荧光素酶活性。总之，肾 IRI 诱导依赖于 HIF-1 α 的 miR-98 上调，从而通过靶向 caspase-3 来保护内皮细胞免于凋亡。 (C) 2015 Elsevier Inc. 保留所有权利。
Endothelial dysfunction is one of the main pathophysiological processes involved in renal ischemia reperfusion injury. Our previous microarray study demonstrated that miR-98 was upregulated in the kidney with ischemia reperfusion injury (IRI). The present study was performed to investigate whether miR-98 was involved in the regulation of endothelial apoptosis under hypoxia and re-oxygenation (H/R) conditions. The dynamic changes of miR-98 in mouse IRI kidney and H/R HUVECs was measured. HUVECs were treated with HIF-1 alpha siRNA to investigate the role of HIF-1 alpha on miR-98 expression. The potential target genes of miR-98 were predicted by bioinformatics analyses. HUVECs were transfected with miR-98 mimics or inhibitor to confirm the role of miR-98 on the expression of target genes and hypoxia-induced apoptosis. The target gene was finally confirmed by dual-luciferase reporter assay. Both of IRI and H/R induced significantly up-regulation of miR-98 in the ischemic kidney and hypoxic HUVECs. HIF-1 alpha siRNA remarkably down-regulated the expression of miR-98 in both normal and hypoxic HUVECs. The putative target genes of miR-98 included IL-6, IL-10 and caspase-3. MiR-98 mimics significantly inhibit caspase-3 expression in HUVECs, while anti-miR-98 significantly up-regulated it. But no change of IL-6 and IL-10 levels was observed after miRNA transfection. miR-98 protected HUVECs against apoptosis induced by hypoxia, while anti-miR-98 had the reverse effect. Furthermore, the dual-luciferase reporter assay confirmed that miR-98 decreased the luciferase activity by targeting the 3' untranslated region of caspase-3. In conclusion, Renal IRI induces up-regulation of miR-98 dependent on HIF-1 alpha, which protects endothelial cells against apoptosis by targeting caspase-3. (C) 2015 Elsevier Inc. All rights reserved.