自噬-溶酶体通路在针刺防治帕金森病模型大鼠中的作用研究

The substantia nigra dopamine neurons degeneration and loss of neurons to form slurry in eosinophilic inclusion bodies in Lewy body are the typical pathological features of Parkinson's disease（PD）. Lewy body is the diagnostic feature of PD, and α-synuclein is the main structure of Lewy body component.α-synuclein with cytotoxic effect is usually oligomer and polymer state. Autophagy-Lysosome Pathway is the more efficient clearance method to remove the abnormal accumulation of protein. Macroautophagy and chaperone-mediated autophagy both participate in the degradation of α-synuclein. The abnormal accumulations of α-syn have relationship with the disorders of Autophagy-Lysosome degradation pathway. Based on this idea, we will carry out the research with preparation of PD model by subcutaneous injection of rotenone method, using acupuncture Fengfu, Taichong therapy on the basis of the preliminary work. Starting from the role of Autophagy-Lysosome Pathway in the pathogenesis of PD induced by rotenone to investigate the effect of acupuncture on the two ways of macroautophagy and chaperone-mediated autophagy. The research will clear the Autophagy-Lysosome Pathway foundation role in the pathogenesis of PD, find prevention and cure effect target of DA neurons of substantia nigra injury, so as to reveal the mechanism of acupuncture therapy in the treatment of PD.

帕金森病（PD）典型病理特征为黑质多巴胺（DA）能神经元变性缺失和神经元包浆内嗜酸性包涵体路易小体（LB）的形成，LB是 PD 的诊断特征，α-突触核蛋白（α-syn）则是 LB的主要结构成分。具有细胞毒性作用的α-syn通常表现为寡聚体和多聚体状态，自噬-溶酶体途径是病理状态下清除异常聚集蛋白质更为有效的清除方式。大自噬和分子伴侣介导的自噬都参与 α-syn 的降解，α-syn 的异常积聚与自噬-溶酶体降解通路障碍有关。基于此思路，本课题拟在前期工作的基础上，采用皮下注射鱼藤酮法制备PD模型，运用针刺风府、太冲进行治疗，从自噬-溶酶体通路在鱼藤酮诱导的PD发病机制中的作用着手，研究针刺对大自噬和分子伴侣介导的自噬这两个途径的影响。力图在明确自噬-溶酶体通路在PD发病机制中的作用基础上，寻求针刺防治黑质DA神经元损伤的作用靶点，揭示针刺疗法治疗PD的作用机制。

本研究采用皮下注射鱼藤酮法制备PD模型，运用针刺风府、太冲进行治疗，从自噬-溶酶体通路在鱼藤酮诱导的PD发病机制中的作用着手，探讨针刺对大自噬和分子伴侣介导的自噬这两个途径的影响。结果经过电针治疗干预后，PD模型大鼠黑质内α-syn阳性神经元数量及其蛋白、自噬底物蛋白P62表达显著降低，酪氨酸羟化酶（TH）阳性神经元数量及其蛋白、微管相关膜蛋白轻链3（LC3-Ⅱ）、组蛋白去乙酰化酶 6（HDAC6）、溶酶体相关膜蛋白2a（Lamp2a）、热休克同源蛋白70（Hsc70）表达显著提高，提示电针风府、太冲能促进LC3-Ⅱ、HDAC6、Lamp2a、Hsc70的表达，并降解α-syn和自噬底物蛋白P62，使自噬流顺利完成，有效防止黑质内DA能神经元的损伤，从而延缓PD的病程。电针防治PD的机制可能与针刺能够改善自噬功能障碍，促进PD模型大鼠体内具有细胞毒性作用α-syn的降解有关。研究结果为揭示针刺治疗PD的作用机制开辟了新的思路，同时为针灸临床治疗PD提供了新的理论依据。