天麻活性成分20C基于Pyk2为靶点的抗帕金森病机制研究

Parkinson's Disease (PD) is a common neurodegenerative diseases. DA accumulation in the cytoplasm caused by the damage of DA release is the main reason of PD pathology. Our previous research has shown that the expression of Proline-rich kinase 2 (Pyk2) is downregulated in the striatum, the pathological tissue of PD, compared with the normal condition. Turnover of the expression of Pyk2 could significantly improve the PD syndrom. Moreover, we also found that the phosphorylation of Pyk2 Y402 was required for DA release, which could attenuate the death of dopaminergic neural cell caused by intracelluar accumulation of DA. These results indicated us that Pyk2 might be a new effective target for anti-PD treatment. 20C is a novel bibenzyl compound isolated from Gastrodia elata. Our study has shown that 20C have an obvious meliorative effect against PD model, while it is mechanism is still unclear. Thus, our research aims to absolve the three aspects of tasks below. At first, we will confirm the role of Pyk2 in PD and its mechanism. Next, we’ll construct the conditional knockout mice model, in which Pyk2 will be knock out specifictly in dopaminergic neural cell. We’ll also construct the drug screening model with dual-luciferased reporter gene system of Pyk2 promoter. Finally, with the models above, we will further investigate the anti-PD effect and mechanism of 20C. Our research will not only provide new drug candidate with intellectual property rights, but also establish a theoretical foundation for the PD new drug reasearch targeting Pyk2.

帕金森病（PD）是常见神经退行性疾病，尚无理想的治疗方法。多巴胺（DA）神经元中DA释放障碍导致的DA蓄积是PD发病且加重的主要原因。我们前期研究发现PD主要病变部位纹状体内酪氨酸激酶Pyk2表达明显降低，而调节其表达回复后PD症状得到显著改善；我们进一步发现Pyk2 Y402位磷酸化参与DA释放，改善胞内DA诱发的神经元死亡，提示我们Pyk2有望成为抗PD的新靶点。我们前期发现传统抗PD中药天麻中提取的联苄类化合物20C对PD病理模型有明显的改善作用，但作用机制尚不清楚。本项目拟就以下方面开展研究：1）确定Pyk2在PD中的作用及其机制；2）构建DA神经元特异性敲除Pyk2的小鼠模型及Pyk2启动子报告基因的细胞筛选模型；3）基于上述体系明确20C的抗PD作用及其机制。本项目的实施不仅为开发具有我国自主知识产权的抗PD新药提供优质候选药物，也为基于Pyk2的抗PD新药提供理论依据。

帕金森病（PD）目前尚无理想的治疗方法。众多研究结果表明多巴胺能（DA）神经元的DA释放障碍引起的DA蓄积毒性是PD发病且加重的主要原因。本项目通过三年的研究，已按计划完成申请书中的研究内容，研究表明Pyk2参与DA释放，对其调节可使PD症状得到改善，进一步研究发现Pyk2通过调节囊泡运动参与递质释放的调节分子，并检测了其相互作用蛋白。在此基础上构建了基于Pyk2为靶点的病理模型及抗PD药物筛选体系。在MPTP动物模型及6-OHDA细胞及动物模型确定20C药效的基础上，进一步检测了其对Pyk2药筛及病理模型的影响，发现Pyk2参与20C的抗PD功效, 提示20C可能通过Pyk2调节DA释放，通过促进自身DA的释放，减少人为给药的局限，并降低胞内DA蓄积引起的毒性损伤。综上所述，本研究阐述了Pyk2在PD中的病理意义，并建立了相关模型，确定了20C在PD模型中的治疗效果及作用机制，本项目的实施不仅为开发具有我国自主知识产权的抗PD新药提供优质候选药物，也为基于Pyk2的抗PD新药提供了理论依据。