MicroRNA-146a down-regulation correlates with neuroprotection and targets pro-apoptotic genes in cerebral ischemic injury in vitro
MicroRNA-146a down-regulation correlates with neuroprotection and targets pro-apoptotic genes in cerebral ischemic injury in vitro
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DOI：
10.1016/j.brainres.2016.07.034
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发表时间：
2016-10
期刊：
影响因子：
2.9
通讯作者：
Xia Zhou;S. Su;Shenghua Li;X. Pang;Chunyong Chen;Jin-pin Li;Jingli Liu
中科院分区：
文献类型：
--
作者：
Xia Zhou;S. Su;Shenghua Li;X. Pang;Chunyong Chen;Jin-pin Li;Jingli Liu
microRNAs（miRNAs）是一类短的非编码RNA，负调控靶基因的表达，在脑缺血损伤中发挥重要作用。MiR-146 a与细胞侵袭、转移、免疫、炎症和凋亡密切相关。已有研究表明miR-146 a可通过不同的病理生理过程抑制或促进细胞凋亡。在我们之前的研究中，血液中的miR-146 a在急性缺血性卒中期间下调。然而，miR-146 a与急性脑缺血损伤之间的联系及其机制尚不清楚。本研究旨在探讨miR-146 a及其可能的靶基因在人SK-N-SH细胞缺氧缺糖16 h和再灌注12 h（OGD/R）损伤中的作用。用miR-146 a模拟物或抑制剂转染细胞以改变miR-146 a的表达。OGD/R损伤后SK-N-SH细胞中MiR-146 a表达下调。此外，生物信息学分析和双荧光素酶分析表明，miR-146 a可直接识别促凋亡基因Caspase 7和Bcl-2相关转录因子1（Bclaf 1）的3′-UTR。miR-146 a过表达可有效降低Caspase 7和Bclaf 1的mRNA和蛋白表达，加重OGD/R诱导的细胞凋亡;而miR-146 a下调则具有神经保护作用。总之，我们的研究表明，miR-146 a有助于OGD/R损伤在体外，同时负调控促凋亡基因，Caspase 7和Bclaf 1。这种特殊的机制为miRNA调控网络提供了新的见解。此外，miR-146 a可能为脑缺血损伤提供潜在的治疗方法。
MicroRNAs (miRNAs) are short, non-coding RNAs that negatively regulate target gene expression, and play an important role in cerebral ischemic injury. MiR-146a has been reported to be highly related to cell invasion, metastasis, immunity, inflammation and apoptosis. Previous studies have indicated that miR-146a can either inhibit or promote apoptosis through different pathophysiological processes. In our previous study, miR-146a in the blood was down-regulated during acute ischemic stroke. However, the connection between miR-146a and acute cerebral ischemic injury and the mechanism underlying the connection remain unclear. Here, we aimed to investigate the role of miR-146a and its possible target genes in human SK-N-SH cells subjected to 16 h of oxygen-glucose deprivation and 12 h of reperfusion (OGD/R) injury. Cells were transfected with miR-146a mimic or inhibitor to alter the expression of miR-146a. MiR-146a in the SK-N-SH cells was down-regulated after OGD/R injury. Moreover, bioinformatics analysis and dual luciferase assays demonstrated that miR-146a directly recognized the 3′-UTR of the pro-apoptotic genes, Caspase7 and Bcl-2-associated transcription factor 1 (Bclaf1). Furthermore, miR-146a over-expression effectively decreased the mRNA and protein expression of Caspase7 and Bclaf1, and aggravated OGD/R-induced cell apoptosis; in contrast, miR-146a down-regulation was neuroprotective. In conclusion, our study revealed that miR-146a contributes to OGD/R injury in vitro, while negatively regulating the pro-apoptotic genes, Caspase7 and Bclaf1. This special mechanism provides new insight into miRNA regulatory networks. In addition, miR-146a may offer a potential therapeutic approach to cerebral ischemic injury.