Xiao Xiao, Tan Zhiwen, Jia Min, Zhou Xiaoli, Wu Kemei, Ding Yanbing, Li Wenjing
Department of Encephalopathy, Hubei Provincial Hospital of Traditional Chinese Medicine (Affiliated Hospital of Hubei University of Traditional Chinese Medicine, Hubei Institute of Traditional Chinese Medicine), Wuhan City, Hubei Province, People's Republic of China.
Department of Encephalopathy, The Central Hospital of Enshi Tujia and Miao Autonomous Prefecture, Enshi City, Hubei Province, People's Republic of China.
Neuropsychiatr Dis Treat. 2021 Apr 22;17:1153-1163. doi: 10.2147/NDT.S286778. eCollection 2021.
Parkinson's disease (PD) is a prevalent neurodegenerative disease. Long noncoding RNA small molecule RNA host gene 1 (SNHG1) has been reported to play critical roles in Parkinson's disease (PD) progression. The study aimed to further elucidate the mechanism of SNHG1 in PD pathogenesis.
The levels of SNHG1, miR-125b-5p and mitogen-activated protein kinase 1 (MAPK1) were determined by quantitative real-time polymerase chain reaction (qRT-PCR) or Western blot. Cell viability and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. The activity of Caspase-3 or Caspase-9 was measured using a Caspase-3 or Caspase-9 Assay Kit. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), IL-1β, lactic dehydrogenase (LDH) activity, reactive oxygen species (ROS) generation and superoxide dismutase (SOD) activity were gauged by enzyme-linked immunosorbent assay (ELISA). Dual-luciferase reporter assay was performed to identify the relationship between miR-125b-5p and SNHG1 or MAPK1. The MPTP-induced PD mouse was used as an in vivo model of PD and MPP-treated SK-N-SH and MN9D cells were used as in vitro models of PD.
SNHG1 and MAPK1 were significantly up-regulated while miR-125b-5p was down-regulated in the MPTP-induced PD mouse model and MPP-induced PD cell models. SNHG1 silence or miR-125b-5p overexpression protected against MPP-evoked apoptosis, oxidative stress and inflammation in SK-N-SH and MN9D cells. Moreover, SNHG1 acted as a molecular sponge of miR-125b-5p, and the protective impact of SNHG1 silence on MPP-evoked cell damage was reversed by miR-125b-5p inhibition. Furthermore, MAPK1 was a functional target of miR-125b-5p and its overexpression attenuated the effects of miR-125b-5p restoration in MPP-triggered cell injury. In addition, the behavioral changes in MPTP-induced PD mouse in vivo model were relieved by SNHG1 silence.
SNHG1 knockdown exerted neuroprotective effects in MPP-evoked cytotoxicity through regulating the miR-125b-5p/MAPK1 axis both in human and mouse PD cell models, highlighting a possible target for PD therapy.
帕金森病（PD）是一种常见的神经退行性疾病。据报道，长链非编码RNA小分子RNA宿主基因1（SNHG1）在帕金森病（PD）进展中起关键作用。本研究旨在进一步阐明SNHG1在PD发病机制中的作用机制。
采用定量实时聚合酶链反应（qRT-PCR）或蛋白质免疫印迹法检测SNHG1、miR-125b-5p和丝裂原活化蛋白激酶1（MAPK1）的水平。分别采用细胞计数试剂盒-8（CCK-8）法和流式细胞术评估细胞活力和凋亡情况。使用Caspase-3或Caspase-9检测试剂盒检测Caspase-3或Caspase-9的活性。通过酶联免疫吸附测定（ELISA）检测肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）、IL-1β、乳酸脱氢酶（LDH）活性、活性氧（ROS）生成和超氧化物歧化酶（SOD）活性。进行双荧光素酶报告基因测定以确定miR-125b-5p与SNHG1或MAPK1之间的关系。MPTP诱导的PD小鼠用作PD的体内模型，MPP处理的SK-N-SH和MN9D细胞用作PD的体外模型。
在MPTP诱导的PD小鼠模型和MPP诱导的PD细胞模型中，SNHG1和MAPK1显著上调，而miR-化酶报告基因测定以确定miR-125b-5p与SNHG1或MAPK1之间的关系。MPTP诱导的PD小鼠用作PD的体内模型，MPP处理的SK-N-SH和MN9D细胞用作PD的体外模型。
在MPTP诱导的PD小鼠模型和MPP诱导的PD细胞模型中，SNHG1和MAPK1显著上调，而miR-125b-5p下调。SNHG1沉默或miR-125b-5p过表达可保护SK-N-SH和MN9D细胞免受MPP诱导的凋亡、氧化应激和炎症。此外，SNHG1作为miR-125b-5p的分子海绵，miR-125b-5p抑制可逆转SNHG1沉默对MPP诱导的细胞损伤的保护作用。此外，MAPK1是miR-125b-5p的功能靶点，其过表达减弱了miR-125b-5p恢复对MPP诱导的细胞损伤的影响。此外，SNHG1沉默可缓解MPTP诱导的PD小鼠体内模型的行为变化。
在人和小鼠PD细胞模型中，SNHG1基因敲低通过调节miR-125b-5p/MAPK1轴对MPP诱导的细胞毒性发挥神经保护作用，这突出了一个可能的PD治疗靶点。
