Department of Neurology, Guta District, The First Affiliated Hospital of Jinzhou Medical University, No.2, Section 5, UniversityLiaoning Province, 121000, China.
Neurotox Res. 2021 Dec;39(6):1771-1781. doi: 10.1007/s12640-021-00437-8. Epub 2021 Nov 13.
Parkinson's disease (PD) is a multi-factorial neurodegenerative disease. Long noncoding RNAs (lncRNAs) have been revealed to be involved in the process of PD. Herein, this study aimed to investigate the potential function and mechanism of JHDM1D-AS1 (JHDM1D antisense 1) in PD process. 1-Methyl-4-phenylpyridinium (MPP +)-induced SK-N-SH cells were used to conduct expression and function analyses. Levels of genes and proteins were examined using real-time reverse transcription PCR (RT-qPCR) and Western blot. Cell viability and apoptosis were determined using CCK-8 assay, flow cytometry, and Western blot, respectively. ELISA analysis was performed for the detection of interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. The contents of lactate dehydrogenase (LDH), superoxide dismutase (SOD), and malondialdehyde (MDA) were measured using commercial kits. The direct interactions between miR-134-5p and PIK3R3 (Phosphoinositide-3-Kinase Regulatory Subunit 3) or JHDM1D-AS1 were verified by dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. JHDM1D-AS1 expression was decreased by MPP + in SK-N-SH cells in a dose- or time-dependent manner. Functionally, JHDM1D-AS1 overexpression attenuated MPP + -evoked neuronal apoptosis, inflammation, and oxidative stress. Mechanistically, JHDM1D-AS1 competitively bound to miR-134-5p to upregulate the expression of its target PIK3R3. Rescue experiments suggested that miR-134-5p upregulation reversed the inhibitory effects of JHDM1D-AS1 on MPP + -induced neuronal injury. Moreover, inhibition of miR-134-5p protected neurons against MPP + -induced neuronal apoptosis, inflammation, and oxidative stress, which were abolished by PIK3R3 silencing. JHDM1D-AS1 protected against MPP + -induced neuron injury via miR-134-5p/PIK3R3 axis, suggesting the potential involvement of this axis in PD process.
帕金森病（PD）是一种多因素神经退行性疾病。长链非编码 RNA（lncRNA）已被证明参与 PD 进程。本研究旨在探讨 JHDM1D-AS1（JHDM1D 反义 1）在 PD 进程中的潜在功能和机制。使用 1-甲基-4-苯基吡啶鎓（MPP+）诱导 SK-N-SH 细胞进行表达和功能分析。使用实时逆转录 PCR（RT-qPCR）和 Western blot 检测基因和蛋白水平。分别通过 CCK-8 测定、流式细胞术和 Western blot 测定细胞活力和细胞凋亡。通过 ELISA 分析检测白细胞介素（IL）-1β、IL-6 和肿瘤坏死因子（TNF）-α。使用商业试剂盒测定乳酸脱氢酶（LDH）、超氧化物歧化酶（SOD）和丙二醛（MDA）的含量。通过双荧光素酶报告和 RNA 免疫沉淀（RIP）测定验证 miR-134-5p 与 PIK3R3（磷脂酰肌醇 3-激酶调节亚基 3）或 JHDM1D-AS1 之间的直接相互作用。MPP+以剂量或时间依赖性方式降低 SK-N-SH 细胞中 JHDM1D-AS1 的表达。功能上，JHDM1D-AS1 过表达可减轻 MPP+诱导的神经元凋亡、炎症和氧化应激。机制上，JHDM1D-AS1 竞争性结合 miR-134-5p 以上调其靶基因 PIK3R3 的表达。挽救实验表明，miR-134-5p 的上调逆转了 JHDM1D-AS1 对 MPP+诱导的神经元损伤的抑制作用。此外，miR-134-5p 的抑制可保护神经元免受 MPP+诱导的神经元凋亡、炎症和氧化应激，而 PIK3R3 的沉默则消除了这种保护作用。JHDM1D-AS1 通过 miR-134-5p/PIK3R3 轴保护 MPP+诱导的神经元损伤，表明该轴可能参与 PD 进程。
