The Stroke Unit, The First People's Hospital of Shangqiu, Shangqiu City, 476100, Henan Province, PR China.
Department of Neurosurgery, The First People's Hospital of Shangqiu, Shangqiu City, 476100, Henan Province, PR China.
Neurotoxicology. 2020 May;78:29-35. doi: 10.1016/j.neuro.2020.02.004. Epub 2020 Feb 10.
Long noncoding RNAs (lncRNAs) have been defined as critical regulators of various human diseases. However, the functions of lncRNAs in Parkinson's disease (PD) have not yet been elucidated. In this study, we investigated the role of lncRNA AL049437 in PD and its underlying mechanism.
An in vivo model of PD was established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), while an in vitro model was created using N-methyl-4-phenylpyridinium (MPP+). Gene expression was evaluated using quantitative reverse transcriptase polymerase chain reaction and western blotting. The effects and mechanism of AL049437 in PD were explored using Cell Counting Kit-8 assay, flow cytometry, enzyme-linked immunosorbent assay, and 2',7'-dichlorodihydrofluorescein diacetate fluorescence assay. The interaction between AL049437, miR-205-5p, and mitogen-activated protein kinase 1 (MAPK1) was evaluated using luciferase reporter and RNA pull-down assays.
The expression of AL049437 was upregulated, while that of miR-205-5p was downregulated in MPTP-induced PD mouse model and MPP+-treated SH-SY5Y cells. Silencing of AL049437 mitigated MPP+-induced neurotoxicity in SH-SY5Y cells, as demonstrated by increased cell viability and reduced cell apoptosis. Furthermore, silencing of AL049437 alleviated MPP+-induced neuroinflammation and oxidative stress, as indicated by the reduction in tumor necrosis factor-α and interleukin-6 levels and reactive oxygen species production. In addition, AL049437 was predominantly localized in the cytoplasm of SH-SY5Y cells and functioned as an miR-205-5p sponge. Moreover, MAPK1 was identified as a downstream target of miR-205-5p. Remarkably, the impact of AL049437 silencing on MPP+-induced neuronal damage could be blocked by miR-205-5p inhibition or MAPK1 overexpression.
Knockdown of lncRNA AL049437 mitigates MPP+ -induced neuronal injury in SH-SY5Y cells by regulating the miR-205-5p/MAPK1 axis. Our research reveals a novel regulatory mechanism of AL049437 in PD progression.
长链非编码 RNA（lncRNA）已被定义为各种人类疾病的关键调控因子。然而，lncRNA 在帕金森病（PD）中的功能尚未阐明。在这项研究中，我们研究了 lncRNA AL049437 在 PD 中的作用及其潜在机制。
使用 1-甲基-4-苯基-1,2,3,6-四氢吡啶（MPTP）建立 PD 的体内模型，使用 N-甲基-4-苯基吡啶（MPP+）建立体外模型。使用定量逆转录聚合酶链反应和蛋白质印迹法评估基因表达。使用细胞计数试剂盒-8 测定法、流式细胞术、酶联免疫吸附测定法和 2'，7'-二氯二氢荧光素二乙酸酯荧光测定法研究 AL049437 在 PD 中的作用和机制。使用荧光素酶报告和 RNA 下拉测定法评估 AL049437、miR-205-5p 和丝裂原活化蛋白激酶 1（MAPK1）之间的相互作用。
在 MPTP 诱导的 PD 小鼠模型和 MPP+处理的 SH-SY5Y 细胞中，AL049437 的表达上调，而 miR-205-5p 的表达下调。沉默 AL049437 减轻了 MPP+诱导的 SH-SY5Y 细胞神经毒性，表现为细胞活力增加和细胞凋亡减少。此外，沉默 AL049437 减轻了 MPP+诱导的神经炎症和氧化应激，表现为肿瘤坏死因子-α和白细胞介素-6 水平以及活性氧产生减少。此外，AL049437 主要定位于 SH-SY5Y 细胞的细胞质中，并作为 miR-205-5p 的海绵。此外，MAPK1 被鉴定为 miR-205-5p 的下游靶标。值得注意的是，miR-205-5p 抑制或 MAPK1 过表达可阻断 AL049437 沉默对 MPP+诱导的神经元损伤的影响。
沉默 lncRNA AL049437 通过调节 miR-205-5p/MAPK1 轴减轻 MPP+诱导的 SH-SY5Y 细胞神经元损伤。我们的研究揭示了 AL049437 在 PD 进展中的新调控机制。
