熟地黄有效成分桃叶珊瑚苷靶向REST调控细胞代谢修复帕金森病多巴胺能神经元DNA损伤的机制研究

帕金森病(PD)是以多巴胺能神经元进行性丢失为主要病理特征的神经退行性病，确切病因未明，尚无长效治疗药物。基因表观遗传修饰参与DNA损伤修复，是PD的重要发病机制。前期发现，熟地黄有效成分桃叶珊瑚苷显著改善PD小鼠神经损伤，上调神经元限制性沉默因子REST的表达并促进其入核，而REST正向调控琥珀酸及延胡索酸的代谢酶，但其作用机制尚不清楚。REST是调控表观遗传的重要基因，据此我们假设，桃叶珊瑚苷靶向REST调控细胞代谢，进而重塑染色质，修复DNA损伤，减缓多巴胺能神经元死亡。本项目通过神经行为学、免疫组织化学、分子生物学等技术方法探讨：1)桃叶珊瑚苷能否靶向REST调控细胞代谢；2)REST调控细胞代谢参与表观遗传修饰的分子机制；3)桃叶珊瑚苷如何靶向REST重塑染色质，修复多巴胺能神经元DNA损伤。以期阐明桃叶珊瑚苷保护多巴胺能神经元的分子机制，为熟地的临床应用提供理论基础和实验证据。

Parkinson's disease (PD) is a neurodegenerative disease characterized with progressive loss of dopaminergic neurons. The exact pathogenesis of PD remains unknown and there is no long-term therapeutic drug yet. Epigenetics is one of the important pathogenesis of PD, and plays a critical role in DNA repair. Our previous studies have shown that the active ingredient Aucubin which was deprived from the Radix Rehmanniae Praeparata significantly improved the neural damages of PD mice. Moreover, Aucubin up-regulated the expression of neuronal restrictive silencing factor REST and promoted the translocation into the nucleus. Moreover, REST positively regulates the metabolic enzymes of succinate and fumarate, however the specific mechanism of action remains unclear. REST is a critical transcription factor in epigenetic modification. Therefore, we hypothetically assumed that Aucubin could target REST to regulate neuronal metabolism and epigenetic remodeling, thereby promoting neuronal DNA damage repair and attenuating dopaminergic neuronal death. In this project, neuroethology, immunohistochemistry, molecular biology and other technical methods were used to explore the following questions: 1) Whether Aucubin can regulate neuronal metabolism by targeting REST; 2) The molecular mechanism by which REST regulates epigenetic modification through cell metabolism; 3) How could Aucubin target REST to remodel chromatin and repair DNA damages of dopaminerigic neurons? Thus the molecular mechanisms of the protective effects for dopaminergic neurons by Aucubin will be elucidated in order to provide theoretical basis and experimental evidences for the clinical application of itself and the Radix Rehmanniae Praeparata.