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微小 RNA-24 的上调通过抑制内皮型一氧化氮合酶的表达导致蛛网膜下腔出血后的血管痉挛。

Upregulation of microRNA‑24 causes vasospasm following subarachnoid hemorrhage by suppressing the expression of endothelial nitric oxide synthase.

机构信息

Department of Neurosurgery, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.

Department of Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.

出版信息

Mol Med Rep. 2018 Jul;18(1):1181-1187. doi: 10.3892/mmr.2018.9050. Epub 2018 May 22.

Abstract

MicroRNA (miR)‑24 has been reported to associate with various diseases by acting on different signaling pathways. The present study aimed to elucidate the association between miR‑24 expression levels and vasospasm following subarachnoid hemorrhage (SAH), and its underlying mechanism. An miR online database was searched, identifying endothelial nitric oxide synthase (NOS3) as a potential target gene of miR‑24. A luciferase reporter assay performed to investigate the regulatory association between miR‑24 and NOS3 revealed that miR‑24 bound to the NOS3 3' untranslated region and inhibited NOS3 expression. Reverse transcription‑quantitative polymerase chain reaction and western blot analysis were performed to investigate the miR‑24 and NOS3 expression levels in samples from patients with SAH, and demonstrated a negative correlation between the two. In addition, miR‑24 expression levels were increased in SAH patients with vasospasm compared with those without, whereas the opposite results were observed for NOS3. Vascular smooth muscle cells (VSMCs) transfected with an miR‑24 inhibitor exhibited increased expression levels of NOS3, whereas those transfected with an miR‑24 mimic or NOS3 small interfering RNA exhibited reduced expression levels of NOS3, compared with the control. These results indicated a negative regulatory association between miR‑24 and NOS3. Downregulation of NOS3 may induce vasospasm following SAH, which may be due to the upregualtion of miR‑24 in VSMCs.

摘要

微小 RNA(miR)-24 已被报道通过作用于不同的信号通路与各种疾病相关。本研究旨在阐明蛛网膜下腔出血(SAH)后 miR-24 表达水平与血管痉挛之间的关联及其潜在机制。通过在线 miR 数据库搜索,确定内皮型一氧化氮合酶(NOS3)为 miR-24 的潜在靶基因。进行荧光素酶报告基因检测,以研究 miR-24 与 NOS3 之间的调控关系,结果表明 miR-24 结合到 NOS3 的 3'非翻译区并抑制 NOS3 表达。逆转录-定量聚合酶链反应和 Western blot 分析用于研究 SAH 患者样本中的 miR-24 和 NOS3 表达水平,结果表明两者之间呈负相关。此外,与无血管痉挛的 SAH 患者相比,血管痉挛患者的 miR-24 表达水平增加,而 NOS3 的表达水平则相反。与对照相比,转染 miR-24 抑制剂的血管平滑肌细胞(VSMCs)中 NOS3 的表达水平增加,而转染 miR-24 模拟物或 NOS3 小干扰 RNA 的 VSMCs 中 NOS3 的表达水平降低。这些结果表明 miR-24 和 NOS3 之间存在负调控关系。NOS3 的下调可能会引发 SAH 后的血管痉挛,这可能是由于 VSMCs 中 miR-24 的上调所致。

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