紫外线诱导海参自溶启动机理的研究

项目名称： 紫外线诱导海参自溶启动机理的研究

项目编号： No.31201299

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 水产食品

项目作者： 启航

作者单位： 大连工业大学

项目金额： 25万元

中文摘要： 海参是自溶能力很强的生物，特别是在受到紫外线的照射后，会出现"吐肠、溶解、化皮"的现象，给海参的贮藏与加工带来不便。研究发现紫外线诱导的海参自溶主要是由于存在于海参的组织蛋白酶（L和B）的作用。然而紫外线作为外界刺激因素导致组织蛋白酶释放的原因，组织蛋白酶释放前发生了哪些信号级联反应，目前尚未确定。针对这两个问题，本项目拟从分子水平上揭示紫外线诱导组织蛋白酶释放前发生的分子信号蛋白质表达，并分析海参体内发生的氧化还原反应。首先，应用免疫印迹技术分析紫外线诱导组织蛋白酶释放前发生的分子蛋白表达，确定组织蛋白酶释放的信号途径；其次应用电子自旋共振（ESR）技术确定活性氧（ROS）的产生方式和种类；最后根据荧光显微镜以及流式细胞仪分析试验，确定海参溶酶体的通透性与破裂，从而为揭示海参自溶启动机理奠定理论基础。

中文关键词： 刺参；自溶；ROS；蛋白酶；氧化应激

英文摘要： Sea cucumber is a kind of living things that possess strong autolysis capacity. The phenomenon of "spiting intestine, dissolution, melting skin" could appear especially sea cucumber is under ultraviolet (UV) irradiation. That takes a lot of inconvenience to the storage and processing of sea cucumbers. Previous study indicated that UV-induced sea cucumbers autolysis was mainly due to cathepsin (L and B) which present in the body of sea cucumber. However, there are something that presently have not been determined, such as the reasons of UV light as the external stimuli factors have led to the release of cathepsin, which signaling cascades happen before cathepsin release.To study these two issues, in the present project, we intend to reveal the molecular protein signaling expression at the molecular level before UV induced cathepsin release, then analysis the redox reactions that occurs in the body of sea cucumber. Firstly, molecular protein expression before UV induced cathepsin release will be analyzed using western blotting and determine cathepsin release of signaling pathways, secondly, the produced manner and type of reactive oxygen species (ROS) will be determined by electron spin resonance (ESR) techniques, finally, based on the test of fluorescence microscope and flow cytometry, the lysosomal permeabil

英文关键词： Stichopus japonicas；autolysis；Reactive oxygen speceies；protease；oxidative stress