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 study the effects of bisphenol A on cell proliferation and expression of estrogen/androgen receptor(ERα/AR) in the human endometrial stromal cells(hESCs).MethodshESCs were isolated and cultured in vitro and cell purity was identified when the cultural confluent condition reached 70%-80%.Cultured cells were treated with concentrations of bisphenol A(0,10-8,10-6 and 10-4mol/L) after synchronization by serum-free culture medium.Forty-eight hours after the treatment with bisphenol A,total RNA of the cultured stroma cells were extracted by Trizol and the mRNA expressions of ERα and AR were detected by real-time PCR.In addition,flow cytometry was used to detect the cell cycles.Results hESCs were successfully isolated and cultured in this study.Confirmed by vimentin immunochistochemical staining,cell purity was about 95%.Cell cycles were significantly inhibited by bisphenol A at the concentrations of 10-6 and 10-4 mol/L.The mRNA expression of ERα in cultured stroma cells was up-regulated by bisphenol A at the concentration of 10-4 mol/L(P0.05).The expression of AR was significantly increased by bisphenol A at the concentrations of 10-6 and 10-4mol/L.However both ERα and AR mRNA were observed to have the trend to be inhibited by bisphenol A of 10-8 mol/L.Conclusion The expressions of ERα and AR in cultured hESCs are significantly inhibited by low concentration,but increased by high concentration of bisphenol A,by which bisphenol A influences endometrial functions.