纳米介导siROCK2调控小胶质细胞活化缓解AD的机制研究

There is no effective way to prevent the progression of Alzheimer disease till now. Our previous studies have found that PEG-PEI/siROCK2 applying to the treatment of AD showed good performance in vitro. Further in vivo experiments have found that it can improve the learning and memory ability of AD model mice. However, the possible mechanism is unclear. Therefore, we propose the scientific question: what is the mechanism of PEG-PEI mediated siROCK2 alleviating AD? Our preliminary experiments have indicated that PEG-PEI/siROCK2 could inhibit the activation of microglia in vitro. Further studies show that it can down regulate the expression of NLRP3 and caspase1 in microglia. On the basis of our previous research findings, we propose a scientific hypothesis: PEG-PEI/siROCK2 would regulate autophagy in microglia, therefore inhibiting neuroinflammation mediated by NLRP3, which consequently leading to the alleviation of AD. This project aims to use nanotechnology to mediating siROCK2 by PEG-PEI, which implying to treat cells or implant into APP/PS1 double transgenic mouse model, through behavior, immunofluorescence, immunohistochemistry and molecular biology method to verify the above assumptions. We hope to provide new methods and theoretical guidance for AD by the interdisciplinary innovative treatment.

阿尔茨海默病在临床上尚无有效的治疗方法。我们前期研究发现纳米PEG-PEI介导siROCK2应用AD表现出良好的体外性能，进一步的体内实验发现其可以改善AD模型鼠的学习和记忆能力，然而其作用机制尚不清楚。因此我们提出的科学问题是：PEG-PEI介导siROCK2缓解AD的机制是什么？我们前期体外预实验发现PEG-PEI介导siROCK2可以抑制小胶质细胞的活化，进一步的预实验发现其可下调小胶质细胞的NLRP3，caspase1的表达。因此，依据前期研究成果，我们提出的科学假设是：PEG-PEI介导siROCK2通过调控小胶质细胞自噬抑制NLRP3炎症小体介导的神经炎症缓解AD。本项目拟应用纳米技术将PEG-PEI载体介导siROCK2，处理细胞或植入APP/PS1双转基因模型鼠，通过行为学，免疫荧光，免疫组化及分子生物学等手段验证上述假设，通过多学科交叉创新为AD治疗提供新策略和理论指导。

阿尔茨海默病在临床上尚无有效的治疗方法。我们前期研究发现纳米PEG-PEI介导siROCK2应用阿尔茨海默病表现出良好的体外性能，进一步的体内实验发现其可以改善阿尔茨海默病模型鼠的学习和记忆能力，然而其进一步的作用以及机制尚不清楚。通过该项目的研究，我们明确了PEG-PEI介导siROCK2作用APP/PS1双转基因小鼠后可以改善小鼠的学习和记忆能力，明确了PEG-PEI介导siROCK2对小胶质细胞无明显毒性作用，明确了PEG-PEI介导siROCK2可以抑制小胶质细胞活化后炎症因子IL-1β和IL-18的表达，明确了PEG-PEI介导siROCK2能够抑制了NLRP3炎症体相关蛋白NLRP3，pro-caspase 1和caspase 1的激活，明确了PEG-PEI介导siROCK2可以激活小胶质细胞的自噬水平，明确了PEG-PEI介导siROCK2是通过激活线粒体自噬水平抑制NLRP3的表达从而下调小胶质细胞炎症反应的机制。本项目的研究结果显示我们明确了纳米材料PEG-PEI介导siROCK2调控小胶质细胞活化缓解阿尔茨海默病的机制。通过本项目的研究，证实了纳米材料PEG-PEI介导siROCK2在体内外均具有明显的治疗效果。因为目前阿尔茨海默病在临床没有有效的治疗方法，这种纳米材料介导基因的技术将来有望应用于临床治疗阿尔茨海默病。本项目研究成果已经发表于SCI杂志，并申请相关专利1项，目前已发表标注该基金号相关SCI论文2篇。目前尚有1篇研究成果论文在投稿中。