Validation and bioinformatic analysis of propofol-induced differentially expressed microRNAs in primary cultured neural stem cells
Validation and bioinformatic analysis of propofol-induced differentially expressed microRNAs in primary cultured neural stem cells
复制标题
丙泊酚诱导原代培养神经干细胞差异表达 microRNA 的验证和生物信息学分析
DOI：
10.1016/j.gene.2018.04.046
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发表时间：
2018-07-20
期刊：
影响因子：
3.5
通讯作者：
Tao, Tao
中科院分区：
文献类型：
--
作者：
Li, Yan;Liu, Youtan;Tao, Tao
丙泊酚是一种广泛使用的静脉麻醉剂，先前被认为是一种神经保护剂。然而，最近越来越多的证据表明，它可能会引起神经毒性，尤其是在神经干细胞（NSCs）的发育过程中。在神经发生过程中导致丙泊酚诱导神经毒性的潜在机制，例如涉及微小RNA（miRNAs）的机制，仍然未知。在本研究中，我们在初步筛选中总共鉴定出27种差异表达的miRNAs，并通过实时定量聚合酶链反应（qRT - PCR）验证了6种miRNAs。其中3种miRNAs上调（miR - 377 - 5p、miR - 194 - 3p和miR - 143 - 5p），3种下调（miR - 3583 - 3p、miR - 466b - 5p和miR - 410 - 5p）。经过基因本体论和京都基因与基因组百科全书（KEGG）通路富集分析，富集在γ - 氨基丁酸能突触通路中的Gabbr1、Cacna1b和Gabbr2被选作可能在丙泊酚诱导的神经毒性中起作用的基因。在丙泊酚暴露后上调的miRNAs所靶向的Gabbr1和Cacna1b在信使核糖核酸（mRNA）和蛋白质水平上表达降低。在丙泊酚处理后下调的miRNAs所靶向的Gabbr2在mRNA和蛋白质表达水平上均上调。在丙泊酚暴露后表现出差异表达的两组miRNAs可能以协同方式在神经干细胞发育过程中同时调节多个基因。我们的研究结果可能有助于阐明分子机制，并为丙泊酚诱导的神经毒性提供潜在的治疗靶点。
Propofol, a widely used intravenous anesthetic, was previously considered as a neuroprotective agent. Recently, however, accumulating evidence suggests that it may cause neurotoxicity, especially in the development of neural stem cells (NSCs). The potential mechanisms contributing to propofol-induced neurotoxicity during neurogenesis, such as those involving microRNAs (miRNAs), are still unknown. In this study, a total of 27 differentially expressed miRNAs were identified in our initial screen and 6 miRNAs were validated by qRT-PCR. Three miRNAs were up-regulated (miR-377-5p, miR-194-3p and miR-143-5p), and three were down-regulated (miR-3583-3p, miR-466b-5p and miR-410-5p). Following gene ontology and KEGG pathway enrichment analysis, Gabbr1, Canca1b and Gabbr2, which are enriched in the GABAergic synapse pathway, were selected as genes potentially playing a role in propofol-induced neurotoxicity. Gabbr1 and Cacna1b, which are targeted by miRNAs that are up-regulated following propofol exposure, showed decreased expression at the mRNA and protein levels. Gabbr2, targeted by miRNAs that were down-regulated following treatment with propofol, was up-regulated at both the levels of mRNA and protein expression. The two clusters of miRNAs that show differential expression following propofol exposure may act in a synergistic manner to regulate several genes simultaneously during the development of NSCs. Our results may contribute to clarify the molecular mechanism and provide potential therapeutic targets for propofol induced neurotoxicity.