基于神经组织芯片和动物精细行为捕捉技术对土家药雪里见的神经毒性物质基础及其机制研究

我国的民族药物在有效性方面具有无可争议的特色与优势，尤其在一些疑难重病和慢性病方面效果突出，但目前民族药的安全性研究基础较差，药物毒性物质基础研究极度匮乏，成为制约民族药快速发展的重要瓶颈。本课题选择有名土家药雪里见，首先利用本团队自主研发的多种动物精细行为捕捉技术，建立更加敏感、系统、规范的雪里见神经毒性评价指标体系。然后运用多种色谱技术分离纯化雪里见中的主要化学成分。之后，在体外培养神经细胞，利用水凝胶材料在体外构建三维的神经细胞模型，结合微流控芯片的流道设计实现快速，可信，高效的药物毒性物质筛选。最后，应用已建立的雪里见神经毒性行为评价指标体系进行验证，同时，开展雪里见损伤小鼠神经系统的神经病理与生化基础研究，以及对动物脑神经突触可塑性相关蛋白表达及cAMP/PKA信号通路的影响，最终建立自动化和智能化程度高的民族药神经毒性评价技术平台，为深入开发有毒民族药物奠定坚实基础。

Effectiveness of ethnic drug in China has incontrovertible features and advantages, especially in treating chronic, difficulty and complicated diseases. However, insufficiency of its safety research basis, and deficiency of basic research of drug toxic substances have become a bottleneck to rapid development in ethnic drug. Arisaema rhizomatum, a famous Tujia herb, was chosen in this study. Capture technologies of animal fine behavior, independently researched and developed by our team, were applied to this study, in order to build a sensitive, normative evaluation indicators system for neurotoxicity of Arisaema rhizomatum. Chromatographic techniques will be used to separate and purify the main chemical in Arisaema rhizomatum. Later, nerve cells will be cultivated and hydrogel will be used to build three-dimensional neuronal cell model in vitro. Furthermore, rapid, credible and efficient drug screening of toxicant can achieve by groove of microfluidic chip. Finally, the evaluation indicators system for neurotoxicity of Arisaema rhizomatum will be used in confirmatory experiment. In addition, neuropathology and biochemical basis tests will be involved in experment of nervous system function damage in mice induced by Arisaema rhizomatum. Whatsmore, the effects of Arisaema rhizomatum on plastic protein expression of animal cranial nerve snapsis and cAMP/PKA signaling pathway will be detected. Technology platforms of neurotoxicity assessment of ethnic drug with high level automation and intelligence will be established at last, to lay a solid foundation for deeper development of toxic ethnic drug.