农药咪鲜安雌激素样作用机制体外研究.docVIP

农药咪鲜安雌激素样作用机制体外研究 目录 TOC \o 1-9 \h \z \u 目录 1 正文 1 文1：农药咪鲜安雌激素样作用机制体外研究 1 1 材料与方法 2 13 试验方法 3 2 结果 4 21 咪鲜安对MCF-7细胞增殖的影响 4 23 雌激素受体（ER）结合试验 5 3 讨论 6 文2：淫羊藿提取物的雌激素样作用研究 6 1 材料与方法 7 2 结 果 10 3 讨论 10 参考文摘引言： 13 原创性声明（模板） 14 文章致谢（模板） 14 正文 农药咪鲜安雌激素样作用机制体外研究 文1：农药咪鲜安雌激素样作用机制体外研究 Study on estrogenic activity and mechanism of prochloraz in vitro Abstrat:Objective To examine the estrogenic-like activity of the pesticide prochloraz by tests in vitro and study the possible MCF-7 cells were maintained in RPMI1640 medium containing 10% bovine weeks before the addition of the test compound the cells were traformed into phenol red-free RPMI1640 medium containing 10% charcoal/dextran treated bovine are seven cell groups deal with different treatments:a vehicle control group,a positive group and five proliferation test,MTT test and growth curve were used to assess the effect on the proliferation of MCF-7 cells by prochloraz,and estrogen receptor(ER)binding test was used to analyze the binding ability of prochloraz with ER. Results Prochloraz can induce the proliferation of MCF-7 cells at 10-7～10-5mol/L with dose-effect relatiohip,and this effect is significantly different from vehicle control(P or P).Our study also show that prochloraz can bind with ER and inhibit the bind of125I-E2 with Prochloraz has estrogenic activity which caused by its bind with ER. Key words: prochloraz;estrogenic activity;MCF-7 cell;estrogen receptor 农药咪鲜安(prochloraz)，系咪唑类广谱杀菌剂，Vinggaard等曾报道过咪鲜安的抗雄激素活性〔1〕。本研究曾通过体内试验证明咪鲜安具有雌激素样作用〔2〕。本文在体外实验条件下观察咪鲜安对人乳腺癌细胞株MCF-7细胞增殖的影响及其与雌激素受体(ER)竞争性结合的能力，对其雌激素作用进行探讨，并研究其可能的作用机制。 1 材料与方法 11 细胞培养 人乳腺癌MCF-7细胞在本实验室常规传代培养。 12 受试物与试剂 咪鲜安原药(纯度98%，浙江绍兴东湖生化有限公司)；17-β-雌二醇(E2)(美国Sigma公司)；125I-E2及分离试剂(北京北方生物制品研究所)；MTT(上海华舜生物工程有限公司)；二甲基亚矾(DMSO，天津市化学试剂一厂)；RPMI1640培养基、无酚红RPMI1640培养基产品(美国Hyclone公司)；高融合型细胞培养用小牛血清、去激素小牛血清(上海纬群生物工程有限公司) 13 试验方法 131 细胞培养与观察 MCF-7细胞培养在含10%小牛血清的RPMI1640培养基中，在37℃恒温、5％CO2、100％湿度下培养。用含025％胰酶、002％乙二胺四乙酸(EDTA)的消化液消化传代