从PINK1和Parkin基因调控角度探讨大补阴丸合牵正散对帕金森病脑线粒体质量控制的分子机制

Parkinson's disease (PD) is a common progressive neurodegenerative disorder. Mitochondrial dysfunction is recognized as the main pathogenesis of PD. Recent studies found that PINK1 / Parkin gene play an important role in mitochondrial quality control and mutation of the two can lead to PD. In Traditional Chinese medicine (TCM), "yin deficiency of liver and kidney" is believed as the key pathogenesis of PD (Vibration Syndrome) and mitochondrial dysfunction can be incorporated into the category of "yin deficiency of liver and kidney ". In the completed project supported by a grant from NSFC, our studies have demonstrated that neuroprotective effects of DBYW and QZS on regulating brain mitochondrial genes expression in mouse models of PD. Based on this work, we proposed to establish the cell model and mouse model of PD by treating with genic modification (overexpression and knockdown of PINK1/Parkin gene) and environmental toxin. Then, we want to explore the effect of DBYW and QZS on mitochondrial morphology and function at the molecular level in PD models. The study aims to explore the molecular mechanism and targets of DBYW and QZS on mitochondrial quality control of PD based on regulation of the Pink1/Parkin Gene. It will enrich modern scientific connotation of "deficiency of liver-yin and kidney-yin" of the etiology and pathogenesis of PD in TCM, provide new idea for treating PD by applying formulas and medicinal of "enriching yin and nourishing kidney, calming the liver and suppressing yang，expelling wind and resolving phlegm", and provide scientific evidence for the combination of formulas in TCM.

帕金森病（PD）是常见的神经退行性疾病，线粒体功能障碍是其主要发病机制，近期研究发现PINK1和Parkin基因在PD线粒体质量控制中发挥着重要作用，二者突变可引起PD。中医认为在PD中的线粒体功能障碍可纳入“肝肾阴虚”的范畴，我们研究发现临床治疗PD的滋阴补肾平肝潜阳有效方药大补阴丸（DBYW）合牵正散（QZS）可以通过调控PD动物脑线粒体基因的表达等达到神经保护作用。在此基础上，本项目采用分子神经生物学和基因等技术，建立PINK1和Parkin基因过表达、敲减与神经毒素联合的PD细胞及动物模型，探讨DBYW合QZS对PD脑线粒体形态和功能及分裂融合的影响。旨在从PINK1和Parkin基因调控的角度明确该方药对PD脑线粒体质量控制的分子机制；丰富PD中医病因病机“肝肾阴虚”的现代科学内涵，为临床“滋阴补肾、平肝潜阳、祛风化痰”方药治疗PD提供新见解,为中医合方理论提供实验依据。

帕金森病（PD）是常见的神经退行性疾病，线粒体功能障碍是其主要发病机制。PINK1和Parkin基因在PD线粒体质量控制中发挥着重要作用，二者突变可引起PD。本项目选用临床治疗PD的滋阴补肾平肝潜阳有效方药大补阴丸（DBYW）合牵正散（QZS），采用分子神经生物学和基因等技术，建立PINK1和Parkin基因过表达、敲减与神经毒素联合的PD细胞及动物模型，探讨DBYW合QZS对PD脑线粒体形态和功能及线粒体分裂融合的影响。本研究发现在PINK1和Parkin基因过表达的水平下，DBYW合QZS能明显改善 PD 小鼠的行为学，减少黑质多巴胺能神经元丢失，减轻黑质神经元的损伤，并提高多巴胺及其代谢产物 DOPAC 、HVA 含量；可提高线粒体形态因子并明显改善线粒体形态和活性；提高脑线粒体酶复合物 I的活性、线粒体膜电位及ATP水平；提高线粒体融合蛋白的表达，但降低线粒体分裂蛋白的表达；可提高PINK1和Parkin蛋白的表达水平并降低PINK1和Parkin的线粒体共定位水平。在PINK1和Parkin基因敲减的水平下，DBYW合QZS的治疗效果尚不明显。实验结果表明DBYW合QZS可以通过增加PINK1和Parkin蛋白的表达，进而维持线粒体分裂融合的动态平衡，更好的发挥对线粒体形态和功能的保护作用，说明DBYW合QZS治疗 PD 的作用机制与 PINK1和Parkin有密切的关系，为DBYW合QZS临床治疗 PD 的作用环节和机制研究提供了进一步的实验依据。