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黄毅娜, 程薇波, 徐培渝, 余文三, 张敏, 李强, 刘玉清. 双酚A和对-壬基酚雌激素受体的竞争性结合[J]. 中国公共卫生, 2004, 20(5): 559-561. DOI: 10.11847/zgggws2004-20-05-28
引用本文: 黄毅娜, 程薇波, 徐培渝, 余文三, 张敏, 李强, 刘玉清. 双酚A和对-壬基酚雌激素受体的竞争性结合[J]. 中国公共卫生, 2004, 20(5): 559-561. DOI: 10.11847/zgggws2004-20-05-28
HUANG Yi-na, CHENG Wei-bo, XU Pei-yu, . Estrogen receptor competitive binding assay of bisphenol A and para-nonylphenol[J]. Chinese Journal of Public Health, 2004, 20(5): 559-561. DOI: 10.11847/zgggws2004-20-05-28
Citation: HUANG Yi-na, CHENG Wei-bo, XU Pei-yu, . Estrogen receptor competitive binding assay of bisphenol A and para-nonylphenol[J]. Chinese Journal of Public Health, 2004, 20(5): 559-561. DOI: 10.11847/zgggws2004-20-05-28

双酚A和对-壬基酚雌激素受体的竞争性结合

Estrogen receptor competitive binding assay of bisphenol A and para-nonylphenol

  • 摘要:
      目的   用雌激素受体竞争性结合试验检测双酚A(BPA)、对-壬基酚(p-NP)的雌激素样活性。
      方法   将SD大鼠进行人工卵巢去除术, 饥饿受体14d后提取子宫胞浆雌激素受体(ER), 受体蛋白定量, 作雌二醇(E2)非标记配体饱和分析, 以确定E2的IC50。随后进行BPA和p-NP的竞争性取代实验, 观察BPA, p-NP是否能与3H-E2竞争性结合大鼠子宫胞浆ER。
      结果   E2的IC50是1×10-9mol/L, BPA、p-NP的IC50分别为2.08×10-5mol/L和2.66R10-5mol/L, BPA、p-NP的相对结合亲和力系数(RBA)分别为0.0048和0.0038, BPA与ER的结合能力稍高于p-NP。
      结论   BPA和p-NP结合ER是发挥其雌激素样效应机制中的重要因素。

     

    Abstract:
      Objective   The estrogen-like activity of bisphenol A and p-nonylphenol was examined with the estrogen receptor competitive binding assay.
      Methods   Adulthealthy female SD rat were ovarietomized and sacrificed 2 weeks later. Cytosol estrogen receptor were abstracted from the uteri. Aliquots of 150μg cytosol protein were added with 1μCi/mol 3 HE2 50μl and increasing concentrations of unlabeled E2, BPA and p-NP, the total volume of reponse was 200μl. The mixtures were incubated at 4℃ 16-18 hours. DCC methods removed free labeled-ligand. At last, the radioactivity was measured.
      Results   The percent inhibitory values(IC50)of E2 was 1×10-9 mol/L. BPA and p-NP can inhibit of 3 H-E2 binding to rat uterine cycotol estrogen receptor with an IC50 of 2.08×10-5 mol/L and 2.66×10-5 mol/L, respectively. The relative binding affinity(RBA)of BPA and p-NP were 0.004 8 and 0.003 8, respectively. The binding capacity of BPA to ER was bigger than that of p-NP.
      Conclusion   Binding to ER maybe was the mechanism that BPA and p-N P mimicked estrogen-like activity.

     

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