In recent years, accumulating articles have revealed that long non-coding RNAs (lncRNAs) play crucial roles in ischemic stroke (IS). A previous study found that lncRNA zinc finger antisense 1 (ZFAS1) was down-regulated in IS patients compared with healthy controls. However, the precise function of ZFAS1 in IS and its associated mechanism remain unclear. Cell viability was assessed by cell counting kit-8 (CCK8) assay. Cell apoptosis was analyzed by flow cytometry. Western blot assay and quantitative real-time polymerase chain reaction (qRT-PCR) were conducted to measure protein and RNA expression. The interaction between microRNA-186-5p (miR-186-5p) and ZFAS1 or MCL1 apoptosis regulator, BCL2 family member (MCL1) was confirmed by dual-luciferase reporter assay, RNA-pull down assay and RNA immunoprecipitation (RIP) assay. IS cell model was established through exposing N2a cells to oxygen and glucose deprivation (OGD). OGD exposure restrained the viability and induced the apoptosis of N2a cells. OGD exposure down-regulated the expression of ZFAS1 and up-regulated the level of miR-186-5p in a time-dependent manner. ZFAS1 overexpression alleviated OGD-mediated injury in IS cell model. MiR-186-5p was identified as a direct target of ZFAS1, and OGD-induced injury in IS cell model was attenuated by the silence of miR-186-5p. MiR-186-5p interacted with the 3′ untranslated region (3′UTR) of MCL1 messenger RNA (mRNA). ZFAS1 positively regulated MCL1 mRNA expression by sequestering miR-186-5p in N2a cells. ZFAS1 overexpression-mediated protective effects in IS cell model were partly overturned by the overexpression of miR-186-5p. MCL1 silencing partly counteracted the protective effects mediated by miR-186-5p silencing in IS cell model. In conclusion, ZFAS1 overexpression exerted a protective role in IS cell model to attenuate OGD-induced injury through targeting miR-186-5p/MCL1 axis. ZFAS1/miR-186-5p/MCL1 signaling might be a novel diagnostic marker and promising treatment target for IS patients.

近年来，越来越多的文章揭示了长链非编码RNA（lncRNA）在缺血性脑卒中（IS）中发挥着至关重要的作用。之前的一项研究发现，与健康对照相比，IS 患者的 lncRNA 锌指反义 1 (ZFAS1) 下调。然而，ZFAS1 在 IS 中的确切功能及其相关机制仍不清楚。通过细胞计数试剂盒 8 (CCK8) 测定法评估细胞活力。通过流式细胞术分析细胞凋亡。进行蛋白质印迹分析和定量实时聚合酶链反应 (qRT-PCR) 以测量蛋白质和 RNA 表达。microRNA-186-5p (miR-186-5p) 与 ZFAS1 或 MCL1 凋亡调节因子、BCL2 家族成员 (MCL1) 之间的相互作用通过双荧光素酶报告基因分析、RNA 下拉分析和 RNA 免疫沉淀 (RIP) 分析得到证实。通过将 N2a 细胞暴露于氧气和葡萄糖剥夺 (OGD) 中建立 IS 细胞模型。OGD暴露抑制了N2a细胞的活力并诱导了细胞凋亡。OGD暴露以时间依赖性方式下调ZFAS1的表达并上调miR-186-5p的水平。ZFAS1 过表达减轻了 IS 细胞模型中 OGD 介导的损伤。MiR-186-5p 被确定为 ZFAS1 的直接靶标，并且 OGD 诱导的 IS 细胞模型损伤因 miR-186-5p 的沉默而减弱。MiR-186-5p 与 MCL1 信使 RNA (mRNA) 的 3' 非翻译区 (3'UTR) 相互作用。ZFAS1 通过隔离 N2a 细胞中的 miR-186-5p 正向调节 MCL1 mRNA 表达。IS细胞模型中ZFAS1过表达介导的保护作用部分被miR-186-5p的过表达所推翻。MCL1 沉默部分抵消了 IS 细胞模型中 miR-186-5p 沉默介导的保护作用。总之，ZFAS1 过表达在 IS 细胞模型中发挥保护作用，通过靶向 miR-186-5p/MCL1 轴减轻 OGD 诱导的损伤。ZFAS1/miR-186-5p/MCL1 信号可能是一种新的诊断标志物和有希望的 IS 患者治疗靶点。