3种环境雌激素检测和筛选方法的比较

Comparison among three screening methods for environmental estrogens

介绍了进行环境雌激素检测和筛选的 3种方法 :鱼类卵黄蛋白原诱导实验法、重组基因酵母检测法和酵母双杂交法 ,着重介绍了这 3种方法的基本原理、具体操作、适用条件及应用现状。卵黄蛋白原诱导实验是根据正常雄性体内不会含有卵黄蛋白原 ,环境雌激素能刺激卵生动物肝细胞产生卵黄蛋白原 ,可通过测定动物体内合成的卵黄蛋白原含量来评价其活性。重组基因酵母法是根据环境雌激素能刺激转化酵母的报告基因表达 ,以表达的β- 半乳糖苷酶的量为依据来判断雌激素活性的大小。酵母双杂交法是在重组基因酵母法的基础上 ,引入了辅激活蛋白 ,通过检测雌激素受体和辅激活蛋白之间的相互作用来评价环境雌激素的活性。这 3种方法均具有灵敏、简便、快速、经济的优点 ,同时既可以定性检测又可以定量检测 。

The present author intends to introduce three bioassay methods to screen the environmental estrogen pollutants, that is, vitellogenin induced in fish, recomibinant yeast assay and yeast two-hybrid assay. As is known, more and more synthetic chemicals with estrogenic activity have been reported to bring about harms to human being as well as wildlife in recent years, which make it urgent to find an effective and secure method to remove the environmental estrogens. Generally speaking, there is no vitellogenin in male animals. The vitellogenins can be synthesized in hepatocytes of oviparous animals by an induction of the environmental estrogens. The estrogenic activity of environmental pollutants can be traced by measuring the quantity of vitellogenin. Environmental estrogen pollutants can cause the expression of the reporter gene (encoding the enzyme β-galactosidase) in recomibinant yeast. Thus, it is estimated that estrogenic activity of chemicals can be detected by measuring the quantity of β-galactosidase. Yeast two-hybrid assay is on the base of recomibinant yeast assay. Since the assay contains coactivator, we can evaluate the presence of estrogenic activities of chemicals by measuring the interaction of estrogen receptor with coactivator. Thus, the paper shows that all of these methods are not only qualitative but also quantitative. And these assays are efficient and rapid techniques for screening and quantitatively analyzing the estrogens in the environmental samples.