circRNA_0114605调控miRNA-29b在水通道蛋白4介导的1,2-二氯乙烷中毒性脑水肿中的作用机制研究

1,2-二氯乙烷（1,2-DCE）中毒性脑水肿严重危害我国职业工人健康，防治的关键在于阐明中毒机制和寻找早期生物学标志。水通道蛋白4（AQP4）在脑水肿中发挥核心作用，微小RNA（miRNA）和环状RNA（circRNA）调控AQP4影响1,2-DCE中毒性脑水肿发生发展鲜见报道。本项目在构建1,2-DCE中毒性脑水肿细胞和动物模型的基础上，应用AQP4高表达质粒或CRISPR-Cas9抑制、miR-29b mimic或抑制剂、circRNA_0114605高表达质粒或siRNA、野生型和基因工程小鼠以及不同暴露水平的人群和病例，从正向和反向在细胞、动物和人群三个水平探讨1,2-DCE通过下调circRNA_0114605的表达促进miR-29b对AQP4的负性调控，AQP4的功能改变介导中毒性脑水肿的发生发展，揭示1,2-DCE中毒性脑水肿发生发展的分子机制和发现生物学标志及治疗靶标。

Brain edema caused by 1,2-dichloroethane (1,2-DCE) exposure induces adverse health effects on occupational workers in China. The key to preventing this disease is to elucidate the poisoning mechanisms and to find early biomarkers. Aquaporin 4 (AQP4) plays a central role in brain edema. There are currently few investigative publications on the role of AQP4 and its microRNAs (miRNAs) and circleRNAs (circRNAs) regulation mechanisms in the occurrence and progression of 1,2-DCE-induced brain edema. Our preliminary results have established cell and animal models for brain edema treated with 1,2-DCE and/or its metabolites. Based on these results, we are going to study the role of circRNA_0114605-regulated miRNA-29b on the AQP4 and its function in the 1,2-DCE-induced brain edema on three levels of cell, animal, exposure population and poisoning cases, respectively. First, an in vitro cell model was applied to analyze the pattern of miRNA-29b and circRNA_0114605 regulations on AQP4 and its function in 1,2-DCE-induced brain edema in the presence or absence of high expression plasmid of AQP4 or CRISPR-Cas9 application, the miRNA-29b mimic or inhibitor, high expression plasmid or siRNA of circRNA_0114605, respectively. Then, miRNA-29b and AQP4 wild type and knockout mice were used to confirm the pattern of AQP4 expression and its regulation by miRNA-29b, as well as to explore the key molecular pathways between the miRNA-29b alteration and pathological change in brain edema induced by 1,2-DCE by using proteomic analysis. Finally, an occupational epidemiology study with healthy exposed subjects and poisoning cases was carried out to confirm the results of the in vitro and in vivo studies. This study will shed light on the role of AQP4 and its miRNA and circRNA regulations in 1,2-DCE-induced brain edema. It will also provide scientific evidence on targets for the development of biomarkers and treatment of this occupational hazard.