温肾养肝方调控Grb2/Ras/Erk信号通路干预帕金森病异动症神经元突触重塑的机制研究

Parkinson's disease is a common chronic progressive neurodegenerative disease in middle-aged and elderly people, and the occurrence of dyskinesia in the middle and late stage is almost inevitable. Recent studies have shown that dyskinesia is associated with neuron synaptic plasticity caused by levodopa replacement therapy. Previous clinical studies have showed that the therapeutic effect of Wenshen Yanggan Decoction on patients with Parkinson's disease is clear. It was found that Wenshen Yanggan Decoction could inhibit the activation of Ras/Erk pathway, but the mechanism was unclear. After proteomics screening and verification, we found that Wenshen Yanggan Decoction can significantly down-regulate Grb2. According to the theory of "yang-qi, suppleness and supplementation of tendons" in traditional Chinese medicine, combined with the treatment characteristics of "warm kidney Yang, nourishing the liver blood" and " Warming Yang to extinguish wind " in the Wenshen Yanggan Decoction, We present the scientific hypothesis, that the effect of Wenshen Yanggan Decoction on dyskinesia is due to intervention of neuronal synaptic plasticity by regulating the Grb2/Ras/Erk signaling pathway. To verify this hypothesis, the animal and cell model are studied in this project. The application of in vitro and in vivo gene over-expression and knockdown, signal pathway inhibition, electron microscopy, molecular biological technique and other research methods are used to discuss the potential mechanisms of the intervention of neuronal synaptic plasticity from different aspects. This study can clarify the molecular mechanism of treatment to dyskinesia and provide more theoretical basis for the further application of Wenshen Yanggan Decoction.

帕金森病是中老年人常见的慢性进展性神经系统变性疾病，中晚期患者异动症的发生几乎不可避免。近期研究报道异动症与左旋多巴替代治疗引起的异常神经元突触重塑有关。前期临床研究发现温肾养肝方治疗帕金森病中晚期异动症疗效明确，实验研究发现温肾养肝方可抑制Ras/Erk通路的激活，但机制不详。通过蛋白组学筛选和验证后发现该方可显著下调Grb2蛋白。根据中医“阳气者，柔则养筋”理论，结合温肾养肝方“温肾阳、养肝血”而“温阳熄风”的治疗特点，我们提出温肾养肝方是否通过调控Grb2/Ras/Erk信号通路干预神经元突触重塑，改善异动症状的科学假说。为验证该理论，本项目研究通过动物和细胞模型，应用离体和在体基因过表达和敲减技术、信号通路抑制、电镜显微技术、分子生物学技术等现代研究方法从不同层面探讨该方改善异常突触重塑的可能机制，以期阐明该方治疗异动症的分子机制，为该方的应用开发提供理论依据。

帕金森病是中老年人常见的慢性进展性神经系统变性疾病，中晚期患者异动症的发生几乎不可避免。近期研究报道异动症与左旋多巴替代治疗引起的异常神经元突触重塑有关。前期临床研究发现温肾养肝方治疗帕金森病中晚期异动症疗效明确，实验研究发现温肾养肝方可抑制Ras/Erk通路的激活，但机制不详。通过蛋白组学筛选和验证后我们发现该方可显著下调Grb2蛋白。根据中医“阳气者，柔则养筋”理论，结合温肾养肝方“温肾阳、养肝血”而“温阳熄风”的治疗特点，我们通过体内体外模型，比较应用温肾养肝方后异动症的相关行为学差异及Grb2表达变化，观察温肾养肝方对纹状体Ras/Erk通路的影响；并通过离体基因敲减技术和在体颅内定向基因过表达技术探讨了温肾养肝方通过调控Grb2以抑制Ras/Erk通路的激活，改善异常突触重塑的神经保护机制，揭示了温肾养肝方治疗阳虚风动为核心病机的帕金森病中晚期LID的分子机制，丰富帕金森病中晚期“阳虚风动”的病机理论，并为该方的应用开发提供更多的实验依据。