Center for Neurodegenerative and Neuroimmunologic Diseases, Department of Neurology, Rutgers Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, and.
Center for Advanced Proteomics Research, Rutgers New Jersey Medical School, Newark, New Jersey 07103.
J Neurosci. 2014 Sep 17;34(38):12725-37. doi: 10.1523/JNEUROSCI.0985-14.2014.
Parkinson's disease (PD) is characterized by the progressive loss of dopaminergic neurons in the substantia nigra. Mitochondrial complex I impairment in PD is modeled in vitro by the susceptibility of dopaminergic neurons to the complex I inhibitor 1-methyl-4-phenylpyridinium (MPP+). In the present study, we demonstrate that microRNA-7 (miR-7), which is expressed in tyrosine hydroxylase-positive nigral neurons in mice and humans, protects cells from MPP+-induced toxicity in dopaminergic SH-SY5Y cells, differentiated human neural progenitor ReNcell VM cells, and primary mouse neurons. RelA, a component of nuclear factor-κB (NF-κB), was identified to be downregulated by miR-7 using quantitative proteomic analysis. Through a series of validation experiments, it was confirmed that RelA mRNA is a target of miR-7 and is required for cell death following MPP+ exposure. Further, RelA mediates MPP+-induced suppression of NF-κB activity, which is essential for MPP+-induced cell death. Accordingly, the protective effect of miR-7 is exerted through relieving NF-κB suppression by reducing RelA expression. These findings provide a novel mechanism by which NF-κB suppression, rather than activation, underlies the cell death mechanism following MPP+ toxicity, have implications for the pathogenesis of PD, and suggest miR-7 as a therapeutic target for this disease.
帕金森病(PD)的特征是黑质中多巴胺能神经元的进行性丧失。PD 中线粒体复合物 I 的损伤在体外通过多巴胺能神经元对复合物 I 抑制剂 1-甲基-4-苯基吡啶鎓(MPP+)的敏感性来建模。在本研究中,我们证明了在小鼠和人类酪氨酸羟化酶阳性黑质神经元中表达的 microRNA-7(miR-7)可保护细胞免受 MPP+诱导的多巴胺能 SH-SY5Y 细胞、分化的人神经祖细胞 ReNcell VM 细胞和原代小鼠神经元的毒性。使用定量蛋白质组学分析鉴定出核因子-κB(NF-κB)的组成部分 RelA 被 miR-7 下调。通过一系列验证实验,证实 RelA mRNA 是 miR-7 的靶标,并且是 MPP+暴露后细胞死亡所必需的。此外,RelA 介导 MPP+诱导的 NF-κB 活性抑制,这对于 MPP+诱导的细胞死亡是必需的。因此,miR-7 的保护作用是通过降低 RelA 表达来缓解 NF-κB 抑制来发挥的。这些发现提供了一种新的机制,即 NF-κB 抑制而非激活是 MPP+毒性后细胞死亡机制的基础,这对 PD 的发病机制具有影响,并提示 miR-7 可作为该疾病的治疗靶点。