【作者】 杨淋清； 罗凌凤； 吴德生； 龚春梅； 黄海燕； 周丽； 杨细飞； 刘建军； 庄志雄；

【Author】 LUO Ling-feng~*,YANG Lin-qing,ZHUANG Zhi-xiong,ZHANG Wen-chang.* Department of Public Health,Fujian Medical University,Fujian 350108,China;Key Laboratory of Modern Toxicology of Shenzhen,Shenzhen Centre for Disease Control and Prevention,Shenzhen 518055,China

【机构】 深圳市疾病预防控制中心；

【摘要】 目的探讨双酚A（bisphenol A,BPA）对小鼠胚胎干细胞的毒性特征及OCT4、SOX2基因表达的影响。方法采用小鼠胚胎干细胞系S6,小鼠胚胎干细胞暴露于BPA24h,倒置显微镜下观察细胞的形态变化,CCK-8法检测BPA对细胞增殖的影响,计算半数抑制浓度（IC50）,并确定染毒剂量;分别采用实时荧光定量PCR和Western blot技术检测双酚A作用下OCT4、SOX2在mRNA和蛋白水平表达的变化。结果 BPA对小鼠胚胎干细胞有一定的毒性作用,IC50为4.3×10^-4mol/L;与对照组相比,10^-7mol/L、10^-6mol/L、10^-5mol/L、10^-4mol/L剂量组OCT4的mRNA相对表达水平明显升高,且具有剂量效应关系,而10^-8mol/L剂量组没有明显改变;与对照组相比,只有最高剂量组(10^-4mol/L)SOX2的mRNA相对表达水平有明显生高。OCT4蛋白相对表达水平,与对照组比较,在10^-5mol/L、10^-4 mol/L剂量组明显升高;而各剂量组SOX2蛋白相对表达水平均未发现明显改变。结论 BPA对小鼠胚胎干细胞具有一定的细胞毒性,在一定剂量范围内BPA可以使小鼠胚胎干细胞OCT4基因表达增加,而对SOX2基因无明显影响。更多还原

【Abstract】 Objective To explore the effects of bisphenol A（BPA）exposure on toxicity characteristic and OCT4 and SOX2 genes expression of mouse embryonic stem cells（mESCs） Methods mESCs were cultured,and treated with different doses of BPA for 24 hours.The morphological changes of mESCs were observed through an inverted microscope.Cell counting kit 8（CCK8） was used to detect the effects of BPA on proliferation of mESCs,and based on the results,the half inhibitory concentration（IC50） was calculated.Real-time fluorescent quantitative polymerase chain reaction（RT-QPCR） and western blotting were used to detect the expression of OCT4 and SOX2. Results BPA had certain toxicity on mESCs,and the IC50 was 4.3×10^-4mol/L.Compared with the control,the OCT4 mRNA expression level increased under the effect of BPA in most groups except in the 10^-8mol/L group.Meanwhile,the SOX2 mRNA expression level increased in the highest exposure group(10^-4 mol/L) of BPA.Moreover,the OCT4 protein expression level of mESCs exposed to 10^-5 mol/L and 10^-4 mol/L BPA increased significantly,while no obvious change of the SOX2 protein expression level was detected in all treated groups.Conclusion BPA in a certain dose range could upregulate the expression of OCT4 gene in mouse embryonic stem cells while had no significant effect on the expression of SOX2 gene更多还原