lncRNA及其个体 NR_030777在百草枯和MPTP致神经细胞损害中的作用及机制

Paraquat is widely used herbicide, which is one of the potential environmental risk factors for Parkinson's disease (PD). Long non-coding RNA (lncRNA) alterations may contribute to neurodegeneration. However, the relationship between lncRNA and effects of environmental neurotoxicant is not known. So far it has not been reported at home and abroad. Our preliminary study found that paraquat and MPTP induce alterations in the expression profiles of lncRNA and particular NR_030777 specificly high expressing in the cytoplasm of dopaminergic neuron in midbrain substantia nigra of PD model mice. We plan to apply gene overexpression and knockdown approaches, through both in vivo animal (intact and gene knockout animals) and in vitro cell experiments, to establish the relationship between lncRNA and PD induced by paraquat or MPTP, to reveal the expressing rules of NR_030777, to explore the relationship between NR_030777 and ZFP326 as well as their functions, to preliminary elucidate the neurobiological functions and mechanisms of NR_030777 in PD induced by paraquat or MPTP. This study will reveal for the first time that the role of NR_030777 regulation function in PD induced by toxicnat and provide important novel insights for potential targets for the prevention and treatment of paraquat-induced PD. This project has original innovation, extremely important theory value and significance of public health practice.

百草枯是应用广泛的除草剂,是帕金森病（PD）的潜在环境危险因素之一。长链非编码RNA（lncRNA）改变可能是神经退行性病变的贡献因子, 但其与环境神经毒物作用的关系不明，迄今国内外未见报道。我们前期研究发现，百草枯或MPTP致PD小鼠中脑黑质lncRNA表达谱改变且其个体NR_030777特异高表达于黑质多巴胺能神经元细胞质内。本项目应用基因过表达与表达抑制策略，体内动物（完整动物和基因敲除动物）和体外细胞实验，建立lncRNA与百草枯或MPTP致PD的关系，明确NR_030777的表达规律，探讨 NR_030777与ZFP326之间的关系及它们的功能，初步阐明NR_030777的神经生物学功能及其在百草枯或MPTP致PD中的作用和机制。将首次揭示NR_030777调控功能在毒物致PD中的作用，为PD防治的潜在靶点提供新视角，有源头创新性和极重要的理论价值和实际公共卫生意义。

百草枯（PQ）是应用广泛的除草剂，是帕金森病（PD）的潜在环境危险因素之一。长链非编码RNA（LncRNA）改变可能是包括PD在内的神经退行性病变的贡献因子，但其与环境神经毒物的关系不明。.本项目应用基因过表达与抑制策略，体内动物（野生动物和基因敲除动物）和体外细胞实验，建立LncRNA与PQ或MPTP致PD的关系，明确LncRNA NR_030777的表达规律，探讨 NR_030777与ZFP326之间的关系，初步阐明NR_030777的神经生物学功能及其在PQ或MPTP致PD中的作用和机制。.本项目获得以下重要的创新性发现：（1）PQ或MPTP暴露可特征性改变中脑黑质组织LncRNA表达谱，NR_030777在小鼠多巴胺能神经元细胞质中特征性高表达。NR_030777可调控其同源基因Zfp326及其潜在下游靶基因Cpne5的表达，改变mRNA及蛋白的表达水平并调控mRNA稳定性。（2）体内外实验均表明，PQ损害神经细胞过程中，经ROS诱导NR_030777的表达升高可促进细胞增殖，加速细胞周期并抑制细胞凋亡。（3）NR_030777参与调控PQ暴露所致的N2a细胞线粒体自噬过程，过表达NR_030777可促进N2a细胞线粒体自噬升高，线粒体自噬标志蛋白LC3B-II表达升高，p62表达水平降低，NBR1水平降低，ATG12-ATG5复合体增多，这些作用在PQ处理N2a细胞后进一步增强。（4）PQ致神经细胞损害中，Zfp326通过与NR_030777结合的SF3B3剪接因子以及影响NR_030777稳定性来调控NR_030777，起到加剧神经毒性的损害作用。（5）炎性LncRNA-AK039862通过Pafah1b1/Foxa1途径调节PQ所抑制的小鼠小胶质细胞和神经细胞的增殖和迁移。这些研究成果首次证明了NR_030777保护神经细胞免受毒物PQ的损害作用，阐明了NR_030777及其同源基因Zfp326之间的调控关系及其在PQ致神经退行性损害中的作用，有明显创新性、极重要的理论价值和实际公共卫生意义。