Department of Neurosurgery, XiangYang Center Hospital, Xiangyang, China.
Department of Neurosurgery, Affiliated Hospital of Hubei University of Arts and Sciences, Xiangyang, China.
J Biochem Mol Toxicol. 2021 Oct;35(10):e22867. doi: 10.1002/jbt.22867. Epub 2021 Aug 8.
Accumulated evidence has manifested that long noncoding RNA (lncRNA) is involved in the progress of Parkinson's disease (PD). SNHG7, a novel lncRNA, has been found to be involved in tumorigenesis. However, SNHG7 expression and its functional effects on PD remain uncharted. Rotenone (Rot) was adopted to construct PD models in Sprague-Dawley (SD) rats and SH-SY5Y cells, respectively. The expression levels of caspase 3, tyrosine hydroxylase (TH), ionized calcium-binding adapter molecule 1 (Iba1) in SD rat striatum were measured via immunohistochemistry and western blot. Additionally, the expressions of inflammatory cytokines (interleukin 1β [IL-1β], IL-6, tumor necrosis factor α) and oxidative stress factors (malondialdehyde, superoxide dismutase, and glutathione peroxidase) in the brain tissues were examined using real-time polymerase chain reaction and enzyme-linked immunosorbent assay, respectively. Moreover, the protein levels of tumor necrosis factor receptor-associated factor (TRAF5), I-κB, nuclear factor-κB (NF-κB), HO-1, Nrf2 were detected via western blot. Bioinformatics was applied to predict the targeting relationship between SNHG7, miR-425-5p, and TRAF5. Dual-luciferase activity assay and RNA immunoprecipitation assays were conducted to verify their interactions. In comparison to healthy donors, SNHG7 was found upregulated while miR-425-5p expression was downregulated in PD patients. Functional experiments confirmed that SNHG7 downregulation or miR-425-5p overexpression attenuated neuronal apoptosis in the Rot-mediated PD model, TH-positive cell loss, and microglial activation by mitigating inflammation and oxidative stress. Mechanistically, SNHG7 served as a competitive endogenous RNA by sponging miR-425-5p and promoted TRAF5 mediated inflammation and oxidative stress. Inhibition of SNHG7 ameliorated neuronal apoptosis in PD through relieving miR-425-5p/TRAF5/NF-κB signaling pathway modulated inflammation and oxidative stress, and similar results were observed in the Rot-mediated rat model of PD.
已有大量证据表明，长非编码 RNA（lncRNA）参与了帕金森病（PD）的进展。SNHG7 是一种新型 lncRNA，已被发现参与了肿瘤的发生。然而，SNHG7 的表达及其对 PD 的功能影响仍未被探明。本研究分别采用鱼藤酮（Rot）构建 PD 大鼠模型和 SH-SY5Y 细胞模型，通过免疫组织化学和 Western blot 检测大鼠纹状体中海马 SNHG7 表达水平，同时检测 caspase 3、酪氨酸羟化酶（TH）、离子钙结合衔接分子 1（Iba1）的蛋白表达。此外，采用实时聚合酶链反应和酶联免疫吸附试验分别检测脑内炎症因子（白细胞介素 1β[IL-1β]、IL-6、肿瘤坏死因子α）和氧化应激因子（丙二醛、超氧化物歧化酶、谷胱甘肽过氧化物酶）的表达。Western blot 检测肿瘤坏死因子受体相关因子（TRAF5）、I-κB、核因子-κB（NF-κB）、血红素加氧酶 1（HO-1）、核因子红细胞 2 相关因子 2（Nrf2）的蛋白表达。采用生物信息学预测 SNHG7、miR-425-5p 与 TRAF5 的靶向关系，通过双荧光素酶活性试验和 RNA 免疫沉淀试验验证它们之间的相互作用。与健康供体相比，PD 患者 SNHG7 表达上调，miR-425-5p 表达下调。功能实验证实，在 Rot 诱导的 PD 模型中，下调 SNHG7 或过表达 miR-425-5p 可通过减轻炎症和氧化应激，抑制神经元凋亡、减少 TH 阳性细胞丢失和小胶质细胞激活。机制上，SNHG7 通过海绵吸附 miR-425-5p 作为竞争性内源性 RNA，促进 TRAF5 介导的炎症和氧化应激。抑制 SNHG7 通过缓解 miR-425-5p/TRAF5/NF-κB 信号通路调节炎症和氧化应激，改善 PD 中的神经元凋亡，在 Rot 诱导的 PD 大鼠模型中也观察到了类似结果。
