重金属离子胁迫下花斑裸鲤钙调蛋白磷酸酶(Calcineurin)的应答及其分子调节机理研究

项目名称： 重金属离子胁迫下花斑裸鲤钙调蛋白磷酸酶(Calcineurin)的应答及其分子调节机理研究

项目编号： No.31460700

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 李长忠

作者单位： 青海大学

项目金额： 50万元

中文摘要： 花斑裸鲤(Gymnocypris eckloni)是黄河上游特有鱼种，是重要的水生野生动物种质资源，也是青海省重要的经济鱼类。近十年的黄河上游水域水质和鱼类资源监测结果显示重金属污染的问题不可忽视。为研究重金属离子胁迫条件下花斑裸鲤应答和分子调节机制，将以铜、锌、铅、镉、汞等为金属污染源，以花斑裸鲤钙调蛋白磷酸酶（Calcineurin，CN）在mRNA、蛋白和酶活性水平的变化确立CN与重金属胁迫间的相关性；以CN在被CsA抑制和非抑制条件下金属硫蛋白（Metallothionein，MT）表达量确立重金属胁迫下CN与MT间的相关性；找到CN和MT应答敏感组织；通过检测CN下游靶基因（IL-1、IL-6和TNF-α）表达量，分析CN及其信号通路的调节机制。探讨建立CN、MT及细胞因子等生物标志物作为重金属污染早期预警体系的可行性，这对制定预防性的管理措施、监控环境污染和资源保护恢复意义重大

中文关键词： 钙调蛋白磷酸酶；花斑裸鲤；重金属；生物标记物

英文摘要： Gymnocypris eckloni, the naked carp species distributed in the Upper Yellow River,is an important wildlife fish germ plasm resource and a significant commercial fish resource of Qinghai province. The result of water quality survey of the Upper Yellow River and monitoring of fish resource in the past 10 years showed that the pollution of heavy metal is worthy of attention. In order to investigate the inner physiological response and molecular regulating mechanism of Gymnocypris eckloni under the stress of heavy metals including copper, zinc, lead, chromium and mercury, the response of Calcineurin (CN) to heavy metal stress will be detected using Real Time PCR, western blotting and enzyme activity determination methods. The responses of Metallothionein(MT) to heavy metal stress will be detected under the condition of CN inhibited or not by CsA, and the correlation between CN and MT responding to heavy metal stress will be established. The responding sensitive tissues of CN and MT to heavy metal stress will be found. The responses of some correlated immunological responsive factors including IL-1,IL-6,TNF-α(the targetd genes of CN)to metal stress will be assayed. The correlation among the heavy metal, CN, MT and immunological responsive factors will be analyzed. The molecular regulating mechanism of CN and its signals transductions pathway responding to heavy metal stress will be primarily elucidated. In this research, the feasibility of making CN, MT and immunological responsive factors as biomarker for predicting the pollution of heavy metal will be probed into, which will play an important role in formulating preventive measures, monitoring of environment pollution, the conservation and restoration of endemic fish species in the Upper Yellow River.

英文关键词： Calcineurin;Gymnocypris eckloni;Heavy metal;Biomarker