Department of Rehabilitation Medicine.
Department of Neurosurgery, Qujing No. 1 Hospital, Qujing, Yunnan, China.
Neuroreport. 2022 Jan 5;33(1):1-12. doi: 10.1097/WNR.0000000000001741.
MicroRNAs are dysregulated in traumatic brain injury and are involved in neuronal cell behaviors. Previous studies identified miR-31 as a spinal cord injury-related microRNA, while its role in traumatic brain injury remains indistinct. Herein, we explored the participation of miR-31 in traumatic brain injury. Traumatic brain injury model was established after traumatic neuron injury. Neurocytes were transfected with miR-31 mimic or inhibitor. Cell counting kit-8, lactate dehydrogenase assay, terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling, and western blot were applied to examine cell viability, lactate dehydrogenase releasing, apoptosis, and apoptosis-related protein. The binding between miR-31 and hypoxia-inducible factor-1A was verified by luciferase assay. Quantitative reverse transcription-PCR was used to detect the regulation of traumatic neuron injury or hypoxia-inducible factor-1A overexpression on vascular endothelial growth factor A level. The effects of hypoxia-inducible factor-1A or vascular endothelial growth factor A on neuronal cell injury were examined. Additionally, phosphatidylinositol 3kinase/AKT/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway was also examined using western blot. Downregulation of miR-31 promoted traumatic neuron injury-induced neuronal cell injury, and its overexpression did the opposite. Hypoxia-inducible factor-1A acted as a downstream mRNA of miR-31 and its downregulation was involved in miR-31-regulated neuronal cell injury. Vascular endothelial growth factor A level was elevated by traumatic neuron injury or hypoxia-inducible factor-1A overexpression. Hypoxia-inducible factor-1A enhanced neuronal cell injury via promoting vascular endothelial growth factor A expression. Furthermore, miR-31/hypoxia-inducible factor-1A/vascular endothelial growth factor A regulated PI3K/AKT/mTOR pathway in neuronal cells. Our study demonstrated miR-31 inhibited neuronal cell apoptosis via regulating hypoxia-inducible factor-1A/vascular endothelial growth factor A axis.
微小 RNA 在创伤性脑损伤中失调,并参与神经元细胞行为。先前的研究确定 miR-31 是一种与脊髓损伤相关的 microRNA,但其在创伤性脑损伤中的作用仍不清楚。本文探讨了 miR-31 在创伤性脑损伤中的参与作用。创伤性神经元损伤后建立创伤性脑损伤模型。用 miR-31 模拟物或抑制剂转染神经元细胞。细胞计数试剂盒-8、乳酸脱氢酶测定、末端脱氧核苷酸转移酶介导的 dUTP 缺口末端标记和 Western blot 用于检测细胞活力、乳酸脱氢酶释放、凋亡和凋亡相关蛋白。通过荧光素酶测定验证 miR-31 与缺氧诱导因子-1A 的结合。定量逆转录-PCR 用于检测创伤性神经元损伤或缺氧诱导因子-1A 过表达对血管内皮生长因子 A 水平的调节。检查缺氧诱导因子-1A 或血管内皮生长因子 A 对神经元细胞损伤的影响。此外,还通过 Western blot 检查磷酸肌醇 3 激酶/AKT/哺乳动物雷帕霉素靶蛋白 (PI3K/AKT/mTOR) 通路。下调 miR-31 促进创伤性神经元损伤诱导的神经元细胞损伤,而上调则相反。缺氧诱导因子-1A 是 miR-31 的下游 mRNA,其下调参与了 miR-31 调节的神经元细胞损伤。血管内皮生长因子 A 水平升高由创伤性神经元损伤或缺氧诱导因子-1A 过表达引起。缺氧诱导因子-1A 通过促进血管内皮生长因子 A 的表达增强神经元细胞损伤。此外,miR-31/缺氧诱导因子-1A/血管内皮生长因子 A 在神经元细胞中调节 PI3K/AKT/mTOR 通路。我们的研究表明,miR-31 通过调节缺氧诱导因子-1A/血管内皮生长因子 A 轴抑制神经元细胞凋亡。