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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3 H -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 0 8× 10 -5mol/L和2 6 6R10 -5mol/L ,BPA、p -NP的相对结合亲和力系数 (RBA)分别为 0 0 0 4 8和 0 0 0 38,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
Objective To detect the estrogen-like activity of bisphenol A (BPA) and p-nonylphenol (p-NP) using estrogen receptor competitive binding assay. Methods SD rats were removed by artificial ovariectomy. The uterine cytoplasmic estrogen receptor (ER) and receptor protein were extracted 14 days after starvation, and the estradiol (E2) non-labeled ligand saturation analysis was performed to determine the E2 IC50. Subsequent competitive substitution experiments with BPA and p-NP were performed to determine whether BPA and p-NP could competitively bind with 3 H -E2 to the uterine cytoplasmic ER of rats. Results The IC50 of E2 was 1 × 10 -9 mol / L, the relative binding affinity coefficients of BPA and p-NP for BPA and p-NP were 208 × 10 -5 mol / L and 266R10 -5 mol / L respectively (RBA) were 0 0 0 4 8 and 0 0 0 38 respectively. The binding ability of BPA to ER was slightly higher than that of p-NP. Conclusion The combination of BPA and p-NP with ER is an important factor in exerting its estrogen-like mechanism