十溴联苯醚暴露对人胚胎干细胞分化潜能的影响

Effects of BDE-209 exposure on differentiation capacity of human embryonic stem cells

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 study the effects of decabrominated biphenyl ether（ BDE-209） exposure on differentiation capacity of human embryonic stem cells. Methods Human embryonic stem cell line FY-h ES-10 cells were exposed to the concentrations of 0（ solvent control）,1,10 to 100 nmol/L BDE-209 for 96 h and then were differentiated into embryoid bodies（ EBs） in vitro. The RNA of EBs derived from FY-h ES-10 cells at Day10 was collected. OCT4,three-layer differentiation-related genes（ AFP,SMA and PAX6）and oxidative stress-related genes（ HIF1 a,HIF2a and SOD1） expression was detected by Q-PCR. The contents of SOD was measured by using kit. Results Compared with the solvent control group,the expression levels of OCT4,SMA,HIF1 a and HIF2 a in BDE-209 exposure groups were increased,the expression levels of AFP,PAX6 and SOD1 weredecreased,with statistically significant differences（ P〈0. 05）. With the increase of the BDE-209 dose,the expression levels of OCT4 showed an increasing trend while the expression levels of SOD1 and the activity of the SOD showed a decreasing trend.Conclusion BDE-209 may interfere with differentiation capacity of human embryonic stem cells resulting in developmental toxicity.