基于定量蛋白组学研究CREB转录因子在铜致小鼠认知障碍中的作用机制

当前，环境中重金属污染特征已发生重要改变，铜暴露的健康风险日益增加。前期研究发现，铜暴露与神经退行性疾病有关，是发生认知障碍不容忽视的原因。本项目拟在构建铜暴露致小鼠认知障碍模型，应用ICP-MS技术对小鼠脏器以及体液进行铜暴露组学分析基础上，通过定量蛋白组学技术分析铜暴露致认知障碍蛋白表达谱，筛选可能与认知功能相关的差异蛋白，重点验证CREB转录因子及相关蛋白的级联响应。以海马神经元细胞HT22为靶细胞，在转录水平和翻译水平探讨CREB在铜暴露小鼠脑组织表达的时相性变化，明确CREB在铜致认知障碍的相关作用通路和调控网络；应用原位杂交、原位荧光染色和RNAScope分析技术，分析靶基因mRNA/蛋白在脑组织的表达和空间位置。最终，阐明转录因子CREB响应铜暴露致认知障碍的分子网络及其作用机制，并筛选早期生物标志及干预靶标，为铜神经毒性的早期预防提供科学依据。

Currently, the characteristics of heavy metal pollution in the environment have changed significantly, with increasing health risks of copper exposure. Our preliminary studies have found that copper exposure is related to neurodegenerative diseases, which is the cause of cognitive impairment. This project intends to build a model of cognitive impairment caused by copper exposure in mice, and uses ICP-MS technology to carry out the histochemical analysis of copper exposure in mice organs and body fluids; Using quantitative proteomic technology to analyze the protein expression profile of cognitive impairment caused by copper exposure, screen differential proteins that may be related to cognitive function, and focus on verifying the cascade response of CREB transcription factors and related proteins. HT22, a hippocampal neuron, was used as the target cell to study the temporal changes of CREB expression in the brain of copper exposed mice at the level of transcription and translation and to clarify the related pathway and regulatory network of CREB in copper induced cognitive impairment. The expression and spatial position of target gene mRNA/protein in the brain were analyzed by in situ hybridization, in situ fluorescence staining and RNAscope method. Finally, the molecular network and mechanism of CREB in response to cognitive impairment caused by copper exposure were elucidated, and early biomarkers and intervention targets were screened to provide scientific basis for early prevention of copper neurotoxicity.