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 【关键词】nbsp; 咪鲜安；雌激素样作用；MCF-7细胞；雌激素受体 　　摘要： 目的nbsp; 用体外试验检测农药咪鲜安的雌激素样作用，并对其可能的作用机制进行探讨。方法nbsp; 将人乳腺癌细胞株（MCF-7）细胞在含10%小牛血清的RPM11640培养基中进行半开放式贴壁培养，加受试物前2周，改在含10%去激素小牛血清的无酚红RPMI 1640培养基中培养。设溶剂对照组、阳性对照组及5个不同浓度的咪鲜安样品组，采用MCF-7细胞增殖试验、四甲基偶氮噻唑蓝（MTT）试验和生长曲线观察咪鲜安对MCF-7细胞增殖的影响，并通过雌激素受体（ER）结合试验分析咪鲜安与ER的结合情况，探讨可能的作用机制。结果nbsp; 咪鲜安在10-7～10-5mol/L剂量范围内可诱导MCF-7细胞增殖，差异有统计学意义（Plt;005或Plt;001），且存在剂量-效应关系；咪鲜安能竞争性地与ER结合，从而抑制125I-E2与ER结合。结论nbsp; 咪鲜安具有雌激素样作用，其作用机制在于其竞争性地与ER的结合。 　　关键词：咪鲜安；雌激素样作用；MCF-7细胞；雌激素受体 　　Study on estrogenic activity and mechanism of prochloraz in vitronbsp; 　　Abstrat:Objectivenbsp; To examine the estrogenic-like activity of the pesticide prochloraz by tests in vitro and study the possible mechanism.Methodsnbsp; MCF-7 cells were maintained in RPMI1640 medium containing 10% bovine serum.Two weeks before the addition of the test compound the cells were transformed into phenol red-free RPMI1640 medium containing 10% charcoal/dextran treated bovine serum.There 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. Resultsnbsp; Prochloraz can induce the proliferation of MCF-7 cells at 10-7～10-5mol/L with dose-effect relationship,and this effect is significantly different from vehicle control(Plt;0.05 or Plt;0.01).Our study also show that prochloraz can bind with ER and inhibit the bind of125I-E2 with ER.Conclusionsnbsp; 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。 　　1nbsp; 材料与方法 　　11nbsp; 细