NLRP3炎症小体在帕金森病α-Syn病理中的作用及机制研究

Recently, a study published by《cell》has showed that NLRP3 gene knockout mice protected against MPTP-induced dopaminergic neuron loss. Therefore, inhibiting the NLRP3 inflammasome activation can be used as a control strategy for PD. However, the role of NLRP3 inflammasome activation in the progression of α-synucleinopathy keeps unknown. Our preliminary study found that NLRP3 inflammasome was activated in a time-dependent manner in the brain of hSYNA53T transgenic mice. Based on these findings, we intend to manipulate NLRP3 inflammasome pathway using lentiviral vectors for gene silencing and overexpression to investigate the effects of NLRP3 inflammasome on α-synucleinopathy. Meanwhile, we also investigate the effect of NLRP3 inflammasome on dysfunctional autophagy in hSYNA53Tmice to explore the underlying mechanism. Primary mouse ventral mesencephalic neuron/glia co-cultures were used to investigate the effect of NLRP3 inflammasome activation on the autophagic flux of neuron. The completion of this project will fully elucidate the contribution of NLRP3 inflammasome to the α-synucleinopathy in PD. More importantly, it will provide a theoretical basis to seek potentially pharmacological intervention strategies targeting NLRP3 inflammasome.

近期，《cell》一项研究显示NLRP3基因敲除小鼠减轻MPTP诱导的多巴胺能神经元的缺失，因此抑制NLRP3炎症小体激活可作为帕金森病（PD）的一个防治策略。但是，NLRP3炎症小体激活在PD的αSyn病理形成中的作用尚不清楚。我们预实验发现hSYNA53T转基因小鼠脑内NLRP3炎症小体时间依赖性地激活。本课题拟应用慢病毒为载体的基因沉默和过表达技术对NLRP3炎症小体途径进行干预，首先研究调控NLRP3炎症小体对hSYNA53T转基因小鼠αSyn病理的影响，然后研究调控NLRP3炎症小体对hSYNA53T转基因小鼠神经元自噬障碍的影响；最后，应用中脑神经-胶质细胞混合培养的方法，离体研究胶质细胞内NLRP3炎症小体激活对神经元自噬流的影响。从而全面阐明NLRP3炎症小体在PD的 αSyn病理中的作用和潜在的机制，为在NLRP3炎症小体这一靶点进行药理学干预提供理论依据。

NLRP3炎症小体激活在帕金森病炎性损伤中发挥重要作用，抑制NLRP3炎症小体激活具有多巴胺能神经保护作用。我们前期研究发现NLRP3炎症小体激活程度与a-Syn病理之间具有正相关性，但是两者之间的直接关系和潜在的机制不清楚。我们使用NLRP3基因敲除小鼠制作MPTP诱导的慢性帕金森病小鼠模型直接研究NLRP3敲除对帕金森病a-Syn病理的影响，同时探讨潜在的机制。. 研究结果显示NLRP3基因敲除抑制MPTP诱导的慢性帕金森小鼠中脑部位胶质细胞活化和神经炎症反应，减轻多巴胺能神经元的降解、a-Syn表达和a-Syn的磷酸化，更为重要的是NLRP3基因敲除提高MPTP诱导的慢性帕金森病小鼠中脑部位蛋白酶活性和改善改善自噬障碍。