报告基因方法检测壬基酚、双酚A抗雄激素样活性的体外研究

Detection of Antiandrogenic Effects of Nonylphenol and Bisphenol A in vitro Reporter Gene-based Assays

[目的]利用基于报告基因的中国仓鼠卵巢细胞(CHO细胞)体外抗雄激素检测方法对壬基酚、双酚A的抗雄激素样活性进行研究,为进一步验证该法在检测抗雄激素样物质的可行性和揭示内分泌干扰作用的机制提供依据。[方法]采用本室建立的瞬时转染人雄激素受体表达质粒pSVAR0、雄激素反应元件调控的报告质粒pMMTV-Luc及内对照质粒phRL-SV40的CHO细胞,在给予1×10-7mol/L二氢睾酮(DHT)后给予不同浓度的壬基酚(NP)及双酚A(BPA),检测报告基因表达活性。[结果]浓度均>1×10-6mol/L的NP、BPA可抑制1×10-7mol/LDHT产生的雄激素样作用,两者具有抗雄激素活性,但较雄激素拮抗剂氟他安(FLU)弱,BPA的抑制作用强于NP。[结论]NP、BPA有一定的抗雄激素活性,BPA的抑制作用强于NP。

[ Objective ] To explore the antiandrogenie effects and the mechanisms of NP and BPA by reporter genebased assays. [ Methods ] Chinese ovary cells were co-transfeeted with the human androgen receptor expression vector, mouse mammary turnout virus MMTV-luciferase vector and phRL-SV40 plasmid for internal control, after treatment of the cells for 24 h with 1×10^-7 mol/L dihydrotestosterone（ DHT ） and NP or BPA, the lueiferase activity was tested. [ Results ] 1 × 10^-6 mol/L NP and 1 × 10^-6 mol/L BPA could antagonize the androgenic effects of 1 × 10^-7 mol/L DHT, NP and BPA are potentially antagonists, but their effects are weaker than flutamide. [ Conclusion ] NP and BPA can antagonize the androgenic effects, and BPA is more powerful than NP. The assay we developed is useful for testing the antiandrogenic effects of chemicals.