多溴联苯醚诱导海洋鱼类生殖细胞凋亡的途径及分子机制研究

项目名称： 多溴联苯醚诱导海洋鱼类生殖细胞凋亡的途径及分子机制研究

项目编号： No.41476109

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 天文学、地球科学

项目作者： 王悠

作者单位： 中国海洋大学

项目金额： 96万元

中文摘要： 阐明鱼类生殖过程对全球变化的响应对于揭示近海生物资源持续衰退的生态环境机制具有重要意义，而生殖细胞凋亡则是分析生殖毒性机理的最直接的方法。本研究因此以海洋鱼类生殖细胞研究为切入点，以全球性新型持久性有机污染物多溴联苯醚(PBDEs)为胁迫因子，将体外细胞毒性与模型动物的体内毒性研究相结合，系统分析PBDEs诱导的生殖细胞凋亡及分子作用机制。我们已经发现PBDEs胁迫过程中通常伴随着活性氧（ROS）升高。为此，本研究采用PBDEs单独或与氧化还原剂联用，通过人为干扰细胞内ROS水平，阐明细胞ROS水平和凋亡的因果关系；通过检测MAPK通路中各元素的变化，阐明生殖细胞凋亡过程中ROS介导的信号通路；通过线粒体途径和内质网途径中标志性指标或元素的变化确定细胞凋亡的具体通路；同时结合动物模型的体内毒性研究，进一步确证毒性作用的途径与机制。研究将为揭示PBDEs生殖毒性的分子机制提供依依据。

中文关键词： 生殖细胞；细胞凋亡；活性氧；分子机制；多溴联苯醚

英文摘要： It is of great significance to reveal the responses of marine fish reproduction to the global changes, because it's one of the most beneficial ways to help clarifying the possible ecological mechanism of the coastal biological resources recession. Studies on germ cell apoptosis is suggested to be the most direct way to elucidate the reproductive toxicity. The present research thus choose the marine fish's germ cell as the breakthrough point and apply the new type of persistent organic pollutant- polybrominated diphenyl ethers (PBDEs) as the stressing factor, and a serious experiements are designed under controlled lab conditions to find the possible pathway and molecular mechanism. We have found in the previous studies that PBDEs exposure could induce the reproductive toxicity in marine zooplankton, increase the reactive oxygen species (ROS) level in the organism, lead to domino effect of oxidative stress and subsequently result in immune responses. However, no causation relation between the ROS level increment and the reproductive toxicity has been studied. Therefore,the aim of the present research is to answer the following questions: 1. what's the role of reactive oxygen species(ROS) in apoptosis induction by PBDEs ? That means we want to know the ROS level increment is the reason or result of PBDEs stress? 2. ROS-mediated pathway could be mitochondria pathway, endoplasmic reticulum pathway and the MAPK kinase pathway, and which one is involved germ cell apoptosis induced by PBDEs? Given the questions above mentioned and basing on our preliminary research results, we change the intracellular ROS levels by redox state modulators (NAC, CAT, Tiron and BSO)to find out the causation relation between ROS level and reproductive toxicity induced by PBDEs . To insight the ROS-mediated signal pathway，we examined the expression of stress activated protein kinase (SAPK) phosphor-JNK、phosphor-p38、JNK、p38. To further confirm the PBDEs-induced apoptotic signal pathway, we examined Cyt c, Caspase3, Caspase9, PARP, GRP78, GRP94, CHOP, Caspase7 and Caspase12 by Western Blot. The in vivo toxic analysis in model fish will be performed simultaneously so that more evidences could be applied for evaluating the effectively mechanism of PBDEs' reproductive toxicity. The results of present study would provide research foundations for revealing the effective pathway and molecular mechanism of PBDEs on marine organisms.

英文关键词： germ cell;apoptosis;reactive oxygen species(ROS);molecular mechanism;polybrominated diphenyl ehters (PBDEs)