大豆异黄酮诱导斑马鱼肝细胞合成VTG的作用机理研究

项目名称： 大豆异黄酮诱导斑马鱼肝细胞合成VTG的作用机理研究

项目编号： No.30870250

项目类型： 面上项目

立项/批准年度： 2009

项目学科： 生物科学

项目作者： 王蔚

作者单位： 中国海洋大学

项目金额： 34万元

中文摘要： 鱼类目前面临着水环境及饵料中植物雌激素的双重暴露危险，金雀异黄素是其中含量最为丰富、活性最强的一种。本项目以卵黄原蛋白vtg-1 mRNA作为环境雌激素的生物标志物，研究发现， 金雀异黄素能显著诱导成年雄性斑马鱼肝脏和离体原代培养肝细胞合成vtg-1 mRNA。在活体实验中，VTG 合成的提高伴随着E2含量的显著升高和睾酮含量的显著下降。离体实验条件下排除了内源E2对VTG的诱导作用，研究表明金雀异黄素需要ERβ21644;ERα21516;时存在才能完全介导斑马鱼原代肝细胞合成VTG，其中起主要介导作用的为ERβ65292;单独ERβ33021;部分介导VTG的合成，单独ERα19981;能介导VTG的合成。同时，金雀异黄素还通过提高ERα21644;ERβ#20004;种受体的产生来促进VTG的合成。但过氧化物酶体增生物激活受体PPARs并不参与金雀异黄素对VTG的诱导作用。由此，本研究初步弄清了金雀异黄素促进斑马鱼合成VTG的作用机理，主要途径有二：一是直接通过ERs，尤其是ERβ20171;导VTG的合成；二是促进体内E2的合成与分泌，从而提高了体内VTG在E2诱导下的合成作用。已发表论文5篇，其2篇为SCI收录论文，3篇为国内核心期刊。

中文关键词： 金雀异黄素；斑马鱼；卵黄原蛋白；作用机理

英文摘要： Fish is now facing the dual exposing conditions of phytoestrogens in both water and diet. Genistein is one of the most aboundant and active phytoestrogens. This project select the vitellogenin vtg-1 mRNA as biomarker of exogenous estrogen, the results show that genistein remarkably induce the synthesis of vtg-1 mRNA in vivo and in vitro of liver of male zebrafish. The in vivo induction of vtg-1 mRNA is accompanied by the marked increase of E2 and decrease of testosterone. The in vitro experiment exluded the induction of VTG by endoestrogen, the results show that the induction of vtg-1 mRNA systhesis in zebrafish primary liver cell culture needs the co-work of ERβnd ERαwith the ERβs the main body. ERβlone may partly mediate the synthesis of vtg-1 mRNA, and ERαlone cannot mediate at all. The synthesis of ERαnd ERβre also improved by the genistein addition. However, the other possible target of genistein - peroxisome proliferator- activated receptors (PPARs), did not mediate the induction of VTG by genistein. Thus, we conclude that genistein may induce the VTG synthesis in zebrafish liver through two ways: 1) directly induce the VTG synthesis throuth the mediation of ERs, especially the ERβ2）promote the production of endoestrogen and then promote the VTG production induced by E2. In the research period, 5 papers were published, including 2 on SCI journals and 3 on Chinese journals. And more 4 papaers are under preparation.

英文关键词： genistein; zebrafish; vitellogenin; action mechanism