Department of Neurology, Chongqing University Central Hospital, Chongqing, China.
Department of Neurology, People's Hospital of Fengdu, Fengdu County, 33 Lutang Street, Sanhe Street, HospitalChongqing, 408200, China.
Neurotox Res. 2022 Aug;40(4):913-924. doi: 10.1007/s12640-022-00523-5. Epub 2022 Jun 7.
Alzheimer's disease (AD) is a progressive neurodegenerative disease, characterized by Amyloid-β accumulation-induced neuronal injury. Emerging evidence shows that circular RNA (circRNA) is involved in AD development. The aim of this study was to illustrate the role of circ-HUWE1 in Amyloid-β accumulation-induced neuronal injury. Quantitative real-time PCR (qPCR) or western blot was conducted for the expression analysis of circ-HUWE1, miR-433-3p, and fibroblast growth factor 7 (FGF7). In functional assays, cell viability was determined by CCK-8 assay, and cell apoptosis was examined by flow cytometry assay, the protein levels of apoptosis-related markers, and caspase1 or caspase3 activity. The release of pro-inflammatory factors was monitored by ELISA. The predicted binding relationship between miR-433-3p and circ-HUWE1 or FGF7 was validated by dual-luciferase reporter assay. We discovered that circ-HUWE1 absence alleviated Amyloid-β-induced cell viability degradation, cell apoptosis, and inflammatory responses in SK-N-SH cells. MiR-433-3p was a target of circ-HUWE1, and miR-433-3p inhibition reversed the effects of circ-HUWE1 knockdown. In addition, FGF7 was a downstream target of miR-433-3p whose function could be abolished by FGF7 reintroduction. Circ-HUWE1 positively regulated FGF7 expression via competitively targeting miR-433-3p. Moreover, circ-HUWE1 knockdown activated the WNT signaling pathway in Amyloid-β-treated SK-N-SH cells by targeting the miR-433-3p/FGF7 axis. In conclusion, circ-HUWE1 knockdown alleviates Amyloid-β-induced neuronal injury in SK-N-SH cells via miR-433-3p release-mediated FGF7 depletion.
阿尔茨海默病（AD）是一种进行性神经退行性疾病，其特征是淀粉样蛋白-β积累诱导的神经元损伤。新出现的证据表明，环状 RNA（circRNA）参与 AD 的发展。本研究旨在阐明 circ-HUWE1 在淀粉样蛋白-β积累诱导的神经元损伤中的作用。通过定量实时 PCR（qPCR）或 Western blot 检测 circ-HUWE1、miR-433-3p 和成纤维细胞生长因子 7（FGF7）的表达分析。在功能测定中，通过 CCK-8 测定法测定细胞活力，通过流式细胞术测定细胞凋亡，测定凋亡相关标志物和 caspase1 或 caspase3 活性的蛋白水平。通过 ELISA 监测促炎因子的释放。通过双荧光素酶报告基因实验验证 miR-433-3p 与 circ-HUWE1 或 FGF7 的预测结合关系。我们发现，circ-HUWE1 缺失减轻了 SK-N-SH 细胞中淀粉样蛋白-β诱导的细胞活力下降、细胞凋亡和炎症反应。miR-433-3p 是 circ-HUWE1 的靶标，miR-433-3p 抑制逆转了 circ-HUWE1 敲低的作用。此外，FGF7 是 miR-433-3p 的下游靶标，其功能可通过 FGF7 再引入而被消除。circ-HUWE1 通过竞争性靶向 miR-433-3p 正向调节 FGF7 的表达。此外，circ-HUWE1 敲低通过靶向 miR-433-3p/FGF7 轴激活了淀粉样蛋白-β处理的 SK-N-SH 细胞中的 WNT 信号通路。总之，circ-HUWE1 敲低通过释放 miR-433-3p 介导的 FGF7 耗竭减轻了 SK-N-SH 细胞中淀粉样蛋白-β诱导的神经元损伤。
