Emerging evidence has unraveled the important implications of microRNAs (miRNAs/miRs) in intracerebral hemorrhage (ICH). The aim of the present study was to assess the possible regulatory role of miR-26a-5p in ICH both in vivo and in vitro. ICH model of rats was constructed using stereotactic injection of VII collagenase, and ICH condition of PC-12 cells was stimulated by hemin. Exogenous overexpression of miR-26a-5p was achieved utilizing the transfection with miR-26a-5p agomir or miR-26a-5p mimics. We detected decreased miR-26a-5p and increased RAN binding protein 9 (RANBP9) levels in perihematomal tissues of ICH rats and in PC-12 cells following ICH. While miR-26a-5p overexpression alleviated behavioral deficits and neuronal apoptosis of rats with ICH. Apoptosis-related proteins Bax, Bcl-2 and cleaved caspase-3 in perihematomal region were also measured to further confirm the inhibitory effect of miR-26a-5p on neuronal apoptosis. In ICH models in vitro, we found that miR-26a-5p overexpression significantly decreased hemin-stimulated apoptosis of PC-12 cells. Additionally, RANBP9 knockdown could suppress the apoptosis of PC-12 cells, similar to the effects of PC-12 cells transfected with miR-26a-5p mimics. With dual-luciferase reporter assay, we identified that miR-26a-5p directly targeted RANBP9. In conclusion, exogenous miR-26a-5p alleviated neuronal apoptosis and brain injury partially by targeting RANBP9, and miR-26a-5p/RANBP9 axis may be a potential target for ICH treatment.

越来越多的证据揭示了microRNA（miRNA / miRs）在脑出血（ICH）中的重要意义。本研究的目的是评估miR-26a-5p在体内和体外在ICH中的可能调节作用。立体定向注射VII型胶原酶建立大鼠ICH模型，血红素刺激PC-12细胞的ICH状态。利用miR-26a-5pagomir或miR-26a-5p模拟物转染可实现miR-26a-5p的外源性过表达。我们在ICH大鼠的血肿组织和ICH后的PC-12细胞中检测到miR-26a-5p降低和RAN结合蛋白9（RANBP9）水平升高。而miR-26a-5p的过表达减轻了ICH大鼠的行为缺陷和神经元凋亡。还测量了血肿周围区域中与凋亡相关的蛋白Bax，Bcl-2和裂解的caspase-3，以进一步证实miR-26a-5p对神经元凋亡的抑制作用。在ICH体外模型中，我们发现miR-26a-5p过表达显着降低了血红素刺激的PC-12细胞的凋亡。此外，RANBP9敲低可以抑制PC-12细胞的凋亡，类似于用miR-26a-5p模拟物转染的PC-12细胞的作用。通过双荧光素酶报告基因测定，我们鉴定出miR-26a-5p直接靶向RANBP9。总之，外源性miR-26a-5p通过靶向RANBP9可以部分缓解神经元凋亡和脑损伤，而miR-26a-5p / RANBP9轴可能是ICH治疗的潜在靶标。