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目的观察双酚A对人子宫内膜基质细胞(hESCs)增殖和雌/雄激素受体(ERα/AR)表达的影响。方法体外培养hESCs,在6孔板贴壁达70%～80%后鉴定细胞纯度。采用无血清培养同步化,然后给予不同浓度的双酚A(0、10-8、10-6、10-4mol/L)处理48 h。Trizol裂解细胞提取总RNA,实时荧光定量PCR法测定基质细胞ERα和AR mRNA表达水平;采用流式细胞技术测定细胞周期。结果本文纯化培养方法得到的hESCs,波形蛋白染色阳性率达95%。10-6、10-4mol/L的双酚A对基质细胞增殖周期有抑制作用。10-4mol/L的双酚A使基质细胞ERαmRNA表达显著升高(P<0.05);10-6、10-4mol/L的双酚A也显著升高AR mRNA的表达水平(P<0.05);但是,10-8mol/L的双酚A则有抑制ERα/AR mRNA表达的趋势(P>0.05)。结论低剂量双酚A抑制人子宫内膜ERα/AR表达,而高浓度双酚A则升高ERα/AR表达,进而影响子宫内膜的功能。
Objective To observe the effects of bisphenol A on the proliferation of human endometrial stromal cells (hESCs) and the expression of estrogen / androgen receptor (ERα / AR). Methods hESCs were cultured in vitro, and the cell purity was identified after the 6-well plates adhered to 70% -80%. Synchronous with serum-free culture, and then given different concentrations of bisphenol A (0,10-8,10-6,10-4 mol / L) for 48 h. Trizol lysed cells were used to extract total RNA. The expression of ERα and AR mRNA in stromal cells was detected by real-time fluorescence quantitative PCR. The cell cycle was determined by flow cytometry. Results The purified hESCs obtained by this method showed a positive rate of 95% for vimentin staining. 10-6,10-4 mol / L of bisphenol A can inhibit the proliferation cycle of stromal cells. 10-4mol / L bisphenol A significantly increased ERαmRNA expression in stromal cells (P <0.05); 10-6,10-4mol / L bisphenol A also significantly increased AR mRNA expression (P <0.05) However, 10-8mol / L bisphenol A had the tendency to inhibit the expression of ERα / AR mRNA (P> 0.05). Conclusions Low dose of bisphenol A can inhibit the expression of ERα / AR in human endometrium, whereas high concentration of bisphenol A increases the expression of ERα / AR and thus affect the function of endometrium.