Graduate School, Tianjin Medical University, Tianjin, China.
Department of Neurology, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning, China.
Kaohsiung J Med Sci. 2020 Oct;36(10):786-792. doi: 10.1002/kjm2.12241. Epub 2020 Jun 3.
Parkinson's disease (PD) is a neurological degenerative disorder that is partially induced by inflammation in the neural system. To explore the roles of disordered microRNAs in the development of PD, we screened 10 miRNAs in the brain samples of 15 postmortem PD patients and 10 postmortem healthy controls by qRT-PCR. The direct targets of miRNAs were predicted by informatics tools and further confirmed by dual luciferase assay and immunoblotting. The function of miRNAs in regulating NF-κB/p65 translocation was examined by immunoblotting, and the overactivation of NF-κB signaling was examined by ELISA. The relationship between dysregulated miRNAs and cytokines was analyzed by correlation analysis. Three miRNAs were found to be reduced in the brains of patients with PD. KPNB1, KPNA3, and KPNA4 were identified as direct targets of miR-218, miR-124, and miR-144. Additionally, KPNA3 was identified as a direct target of miR-124, and KPNA4 was a direct target of both miR-124 and miR-218. The p65 translocation from the cytoplasm to the nucleus was repressed by miR-124, miR-218, and miR-144 in the SH-SY5Y cells. The NF-κB signaling pathway was overactivated after miRNA inhibitor transfection. The upregulation of KPNB1, KPNA3, and KPNA4 in the brain samples of PD patients was confirmed by immunoblotting, and negative correlations were found between dysregulated miRNAs and cytokines. In conclusion, we identified that the downregulation of miR-218, miR-124, and miR-144 in the brain was related to PD via activation of NF-κB signaling, helping to unveil the role played by dysregulated miRNAs in the pathogenesis of PD and provide new potential targets for PD treatment.
帕金森病（PD）是一种神经系统退行性疾病，部分由神经系统炎症引起。为了探索失调的 microRNA 在 PD 发展中的作用，我们通过 qRT-PCR 从 15 名 PD 患者和 10 名尸检健康对照者的脑组织样本中筛选了 10 个 miRNA。利用信息学工具预测 miRNA 的直接靶标，并通过双荧光素酶报告基因实验和免疫印迹进一步验证。通过免疫印迹检测 miRNA 调节 NF-κB/p65 易位的功能，通过 ELISA 检测 NF-κB 信号的过度激活。通过相关性分析分析失调 miRNA 与细胞因子之间的关系。发现三种 miRNA 在 PD 患者的大脑中减少。KPNB1、KPNA3 和 KPNA4 被鉴定为 miR-218、miR-124 和 miR-144 的直接靶标。此外，KPNA3 被鉴定为 miR-124 的直接靶标，而 KPNA4 是 miR-124 和 miR-218 的共同直接靶标。miR-124、miR-218 和 miR-144 在 SH-SY5Y 细胞中抑制 p65 从细胞质向细胞核易位。miRNA 抑制剂转染后 NF-κB 信号通路过度激活。通过免疫印迹证实了 PD 患者脑组织样本中 KPNB1、KPNA3 和 KPNA4 的上调，并且失调 miRNA 与细胞因子之间存在负相关。总之，我们发现大脑中 miR-218、miR-124 和 miR-144 的下调与 PD 通过 NF-κB 信号的激活有关，有助于揭示失调 miRNA 在 PD 发病机制中的作用，并为 PD 治疗提供新的潜在靶点。
