cAMP-PKA介导的高糖抑制团头鲂呼吸爆发功能的作用机制研究

项目名称： cAMP-PKA介导的高糖抑制团头鲂呼吸爆发功能的作用机制研究

项目编号： No.31472289

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 戈贤平

作者单位： 中国水产科学研究院淡水渔业研究中心

项目金额： 80万元

中文摘要： 本项目针对高糖导致鱼体呼吸爆发功能障碍的关键问题，以团头鲂为研究对象，采用整体高糖投喂动物实验和离体高葡萄糖胁迫鱼体白细胞的手段，运用细胞培养、流式细胞仪、荧光定量PCR和Western bolting以及激光共聚焦显微镜、电子显微镜等方法，采用G6PD抑制剂阻断G6PD代谢途径，重点阐明G6PD、NOX、ROS、SOD、MPO与高糖抑制团头鲂呼吸爆发功能的关系，解析高糖导致团头鲂呼吸爆发功能障碍除抑制G6PD活性外，是否还存在其他抑制机制。在此基础上，采用PKA特异性抑制剂PKI和激活剂Forskolin阻断cAMP-PKA信号通路，探索高糖导致团头鲂呼吸爆发功能障碍与CAMP-PKA信号转导通路的关系，进一步阐明cAMP-PKA通路及下游通路在高糖胁迫导致团头鲂呼吸爆发功能障碍的可能信号转导机制，为团头鲂作为草食鱼类却不能利用高糖提供证据，也为高糖影响团头鲂非特异免疫提供理论基础。

中文关键词： 高糖；呼吸爆发功能；信号通路；非特异免疫；团头鲂

英文摘要： Our objective of study is to carry out the study of dietary high carbohydrate on molecular signal transduction mechanism for respiratory burst dysfunction for fish based on Wuchang bream feeding with high carbohydrate and white cells in vitro under high glucose content. By using cell culture, flow cytometry, fluorescent quantitative PCR and western method of bolting, and confocal microscopy, electron microscopy technologies, we will check the changes of key enzymes(G6PD, NOX, ROS, SOD,MPO), respiratory burst activity, signal transduction passway (cAMP, ERK, CJNK, p38MAPK, extracelluar signal-regulate kinase 5) and organism structure. In order to illustrate the relationship between G6PD, NOX, cAMP/PKA and respiratory burst dysfunction, to further explore the signal mechanism for respiratory burst dysfunction induced by dietary high carbohydrate, we use the G6PD specific inhibitor (Dehydrooepian-drosterane)in the metabolism of G6PD, PKA stimulator(Forskokin) and specific PKA inhibitor (PKI) in the pathway of cAMP-PKA in the experiment. We expect to clarify signal mechanism for respiratory burst dysfunction induced by dietary high carbohydrate of Wuchang bream mediated by cAMP-PKA, to reveal some reasons of lower carbohydrate requirement for Wuchang bream as herbivorous animal, and to offer some research ideas of non-specific immunity of high carbohydrate for the fish.The results will provide a theroetical basis for the development efficient feed and health for Wuchang bream.

英文关键词： High carbohydrate;Function of respiratory burst;Signal passway of cAMP/PKA;Non-specific immunlity;Megalobrama amblycephala