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目的探讨十溴联苯醚(BDE-209)暴露对人胚胎干细胞分化潜能的影响。方法分别将对数生长期未分化的人胚胎干细胞FY-h ES-10细胞暴露于含有终浓度为0(溶剂对照)、1、10和100 nmol/L BDE-209的培养基中培养96 h后,采用悬浮法培养拟胚体(embryoid body,EB)自发分化10 d,实时荧光定量PCR检测EB中OCT4、内胚层标志基因AFP、外胚层标志基因PAX6、中胚层标志基因SMA及氧化应激相关基因HIF1a、HIF2a和SOD1基因的表达,并测定SOD活性。结果与对照组比较,各剂量BDE-209暴露组FY-h ES-10细胞EB分化10 d OCT4、SMA、HIF1a和HIF2a表达水平均较高,而AFP、PAX6、SOD1表达水平均较低,差异有统计学意义(P<0.05)。且随着BDE-209暴露剂量的升高,FY-h ES-10细胞分化10 d EB中OCT4的表达水平均呈上升趋势,SOD1基因的表达水平呈下降趋势。同时BDE-209暴露后FY-h ES-10细胞分化10 d EB中SOD活性降低。结论 BDE-209可能通过影响早期胚胎的分化潜能从而产生发育毒性。
Objective To investigate the effects of decabromodiphenyl ether (BDE-209) on the differentiation potential of human embryonic stem cells. Methods FHD-h ES-10 cells were cultured in culture medium containing BDE-209 at final concentration of 0 (solvent control), 1, 10 and 100 nmol / L for 96 h Then the embryoid body (EB) was cultured for 10 days by suspension method. OCT4, AFP, PAX6, mesoderm marker gene SMA and oxidative stress were detected by real-time fluorescence quantitative PCR Related genes HIF1a, HIF2a and SOD1 gene expression, and determination of SOD activity. Results Compared with the control group, the expression levels of OCT4, SMA, HIF1a and HIF2a in EBV-treated ESBLs-1 and EB-1 cells at each dose of BDE-209 exposure group were higher than those in the control group, while the expression levels of AFP, PAX6 and SOD1 were lower There was statistical significance (P <0.05). And with the increase of BDE-209 dose, the expression of OCT4 in FY-h ES-10 cells tended to increase and the expression level of SOD1 decreased. At the same time, the SOD activity of FY-h ES-10 cells differentiated for 10 d after BDE-209 exposure was decreased. Conclusion BDE-209 may cause developmental toxicity by affecting the differentiation potential of early embryos.