本项目以PD小鼠和体外PC12细胞为实验模型，观察Nrf2/ARE通路及其预先激活对百草枯损害黑质神经元和PC12细胞中的保护作用；应用特异性药理学试剂从泛素蛋白酶体功能等来研究百草枯激活Nrf2的机制。本项目首次发现，PQ处理小鼠和PC12细胞可激活神经细胞Nrf2/HO-1途径，tBHQ预处理可削弱百草枯所致的神经退行性病变，可能经过Nrf2/ARE-HO-1途径的激活发挥保护作用；PQ致神经细胞Nrf2核转位（激活）的途径不涉及泛素蛋白酶体降解Nrf2途径和Nrf2 mRNA的诱导，而涉及Nrf2蛋白合成途径。首次发现，蛋白酶体20s复合物胰蛋白酶活性的抑制、ROS生成增加、巯基含量变化可能是PQ致神经细胞损害的机制之一。首次发现，PQ或MPTP致神经退行性病变时呈现miRNA表达的变化，而且Nrf2与miRNA之间存在网络联系，Nrf2在PQ或MPTP致miRNA表达变化中可能起一定作用。此外，还成功构建了Nrf2的microRNA表达载体pcDNA-Nrf2-C和pARE-luc报告基因质粒。研究发现具有极其重要的理论和公共卫生意义。

The project is to explore the protective role of nuclear factor erythroid 2-related factor (Nrf2)-antioxidant responsive element (ARE) pathway and it's pre-activation for damage of neural cells in the substantia nigra and PC12 cells caused by paraquat, using the experimental model of Parkinson's disease (PD) in mice and PC12 cells in vitro. The project is also to investigate the mechanism of Nrf2 activation by paraquat from the ubiquitin-proteasome function, application specific pharmacology reagents. The project for the first time found that the Nrf2-heme oxygenase 1 (HO-1) pathway of neural cells in mice and PC12 cells was activated by PQ and pretreatment with tert-butylhydroquinone (tBHQ) conferred neuroprotection against PQ-induced neurodegeneration probably by increasing Nrf2 and HO-1 expression. Nrf2 activation or nuclear translocation in neural cells by PQ is associated with Nrf2 protein biosynthesis (translation) pathway, not ubiquitin-proteasome degradation pathway and mRNA induction of Nrf2. The project for the first time found that depression of 20s proteasome trypsin activity, increase of ROS generation and change of thiol level is possibly one of mechanisms underlying neural cells' damage caused by PQ. The project for the first time found that miRNA expression was changed in neurodegeneration induced by PQ or 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Nrf2 was correlated with microRNA (miRNA) in network relationship. Nrf2 may play a role in the change of miRNA expression induced by PQ or MPTP. In addition, the miRNA eukaryotic expression vector of Nrf2 (pcDNA-Nrf2-C) and reporter gene plasmid have been successfully constructed. These founding are extremely important theoretical and public health significance.

本项目以PD小鼠和体外PC12细胞为实验模型，观察Nrf2/ARE通路及其预先激活对百草枯损害黑质神经元和PC12细胞中的保护作用；应用特异性药理学试剂从泛素蛋白酶体功能等来研究百草枯激活Nrf2的机制。本项目首次发现，PQ处理小鼠和PC12细胞可激活神经细胞Nrf2/HO-1途径，tBHQ预处理可削弱百草枯所致的神经退行性病变，可能经过Nrf2/ARE-HO-1途径的激活发挥保护作用；PQ致神经细胞Nrf2核转位（激活）的途径不涉及泛素蛋白酶体降解Nrf2途径和Nrf2 mRNA的诱导，而涉及Nrf2蛋白合成途径。首次发现，蛋白酶体20s复合物胰蛋白酶活性的抑制、ROS生成增加、巯基含量变化可能是PQ致神经细胞损害的机制之一。首次发现，PQ或MPTP致神经退行性病变时呈现miRNA表达的变化，而且Nrf2与miRNA之间存在网络联系，Nrf2在PQ或MPTP致miRNA表达变化中可能起一定作用。此外，还成功构建了Nrf2的microRNA表达载体pcDNA-Nrf2-C和pARE-luc报告基因质粒。研究发现具有极其重要的理论和公共卫生意义。