环境雌激素对牙鲆性腺分化的作用和机制的研究-营养与食品卫生学专业论文.docxVIP

复旦大学硕士学位论文 次研究先尝试用E2作为工具药物、运用质粒转染方法，在几种细胞株内傲 P450arom--荧光素酶报告基因分析。结果显示，P450arom转录活性能够被E2显 著诱导，诱导途径是E2与雌激素受体亚型ERa结台而发挥作用，但不是通过ERl3 途径；这一诱导作用只发生在人类卵巢粒层细胞状肿瘤细胞株KGN细胞内(一种 能表达P450arom mRNA而生成类固醇的细胞株)，而不发生在不生成类固醇的 Hepa．E1、COS一7和E293数种细胞株中。据此推测，雌激素类诱导P450arom转 录活性可能是通过存在于KGN细胞中某些特殊因子介导而间接作用的。 最后，为探索另一重要基因MIS在牙鲆幼鱼性腺分化过程中的作用，本试验设 计在18℃雌性化水温中，将重组MIS喂食遗传学雌性牙鲆幼苗，再检测鱼的性别比 例和相关基因的表达。结果显示，喂食重组MIS后，可以诱导18℃水温中部分幼鱼 的表型雄性化，同时用原位杂交检测到变性的表型雄鱼??生殖腺中P450arom mRNA表达受到了抑制，提示MIS基因在鱼的性腺中具有抑带1]P450arom mRNA表 达的作用而改变鱼的性别，即雄性化作用。 本研究的结果提示，环境雌激素对牙鲆性腺分化的作用机制可能是：它们先 与细胞的ERa受体结合诱导P450arom转录，从而提高P450arom表达，抑制MIS表达。 关键词：雌激素样作用 芳香酶P450 苗勒氏管抑制物 性分化 2  复旦大学硕士学位论文 Study on Action Mechanism of Environmental Estrogens on Gonadal Sex Differenfiafion in Flounder(Paralichthys ofivaceus) Abstract Along with the development of industry,the pollution of Environmental Es仃ogens aggravated and became a potential harmful factor to the health ofhuman and wild life．HoweveL their action mechanism on sex differentiation is yet to be clarified． 111e purpose of this study was focused on this question through a series of tests in vivo and in vitro of the flounder(Paralichthys olivaceus)．111is study possessed certain academic significance and practical value on perceiving the hazard，illuminating definite mechanism，enhancing evaluation level and preventing unhealthy effects of Environmental Estrogens．Paralichthys olivaceus is a kind of flounder which belongs to teleost fish that has a chromosome of xx(female)／XY(mal曲sex determination mechanism like human being．111e genetic XX larvae can be experimentally sex—reversed to phenotypic males by rearing the larvae at 27℃water temperature or by treatment with sex steroid hormones and the gonad develops into obvious testis in phenotypic male flounder．Therefore，this kind of fish provides a valuable new animal model for research to elucidate the sex differentiation mechanism influenced by environmental factors． In order to investigate the action and mechanism of Environmental Es