贝类 Nrf2/Keap1 信号通路参与消减微囊藻素毒性的分子机制

Nrf2/Keap1 signaling pathway play an important role in enhancing antioxidant capacity of tissues and protecting the tissues against poison damage etc. Microcystins are an algae toxins, which are generated by Microcystis aeruginosa in the process of the cyanobacterial blooms. Cristaria plicata is one of freshwater pearl bivalves in the aquaculture industry of China. Relatively little is known how Nrf2/Keap1 signaling pathway of mollusks is activated, which regulate transcriptional expression of detoxification and antioxidant enzyme genes at present. Therefore, the project will clone relevant genes of Nrf2/Keap1 signaling pathway, antioxidant target genes and their promoter from C. plicata. The structural feature of these genes will be identified, and the function of promoter will be analyzed. The transcriptional activity of downstream target genes activated by Nrf2、Keap1、Maf and their compound will be verified. The tissue expression of target genes and relevant genes from the signaling pathway, and the expression change after microcystins challenge will be detected. Keap1 interaction with microcystins, and Nrf2, keap1, Maf recombinant proteins combination with ARE sequence will be analyzed. The nuclear translocation of Nrf2 will be observed. The oxidative stress reaction of the signaling pathway will be made clear from microcystins stress in cells. Not only can the information of the project come into being importantly academic values in theory, but also these can provide theoretical foundation for the healthful aquaculture of mollusks in the application.

Nrf2/Keap1信号通路在增强组织抗氧化能力和保护组织免受毒物损伤等方面起着重要的作用。微囊藻毒素是蓝藻水华过程中由铜绿微囊藻产生的毒素。褶纹冠蚌是我国的淡水育珠蚌之一。目前，还不清楚微囊藻毒素如何激活贝类Nrf2/Keap1 信号通路，通路又是怎样调控解毒酶和抗氧化酶基因的转录表达。本项目拟通过克隆褶纹冠蚌Nrf2/Keap1信号通路相关的基因、靶基因及启动子，分析基因的结构特征，鉴定启动子的功能；探明Nrf2、Keap1、Maf及其复合物对下游靶基因的转录活性；检测信号通路相关基因与靶基因的组织表达及微囊藻毒素刺激蚌后的组织表达变化；分析微囊藻毒素与Keap1的相互作用及Nrf2、keap1、Maf重组蛋白与ARE序列的结合；观察Nrf2的核移位情况；探清信号通路对微囊藻毒素胁迫细胞的氧化应激反应。这不仅在理论上具有重要的学术价值，而且在应用上可以为我国贝类的健康养殖提供理论基础。

Nrf2/Keap1信号通路在增强组织抗氧化能力和保护组织免受毒物损伤等方面起着重要的作用。微囊藻毒素是蓝藻水华过程中由铜绿微囊藻产生的毒素。本项目克隆了褶纹冠蚌Nrf2/Keap1信号通路相关的基因、靶基因及启动子，分析基因的结构特征，鉴定了启动子的功能；探明了Nrf2、Keap1、Maf及其复合物对下游靶基因的转录活性；检测了信号通路相关基因与靶基因的组织表达及微囊藻毒素刺激蚌后的组织表达变化；分析了微囊藻毒素与Keap1的相互作用及Nrf2、keap1、Maf重组蛋白与ARE序列的结合。探清信号通路对微囊藻毒素胁迫细胞的氧化应激反应。这不仅在理论上具有重要的学术价值，而且在应用上可以为我国贝类的健康养殖提供理论基础。