双酚A对小鼠胚胎干细胞分化潜能的影响  

Effect of bisphenol A on differentiation potential of mouse embryonic stem cells

作　　者：罗凌凤[1] 崔栋[2] 龚春梅[3] 吴德生[2] 黄海燕[2] 刘建军[2] 张文昌[1] 庄志雄[2] 杨淋清[2] 

机构地区：[1]福建医科大学公共卫生学院环境与健康重点实验室,福建福州350108 [2]深圳市疾病预防控制中心现代毒理学重点实验室深圳市卫生毒理学重点实验室,广东深圳518055 [3]深圳市慢性病防治中心,广东深圳518020

出　　处：《中国药理学与毒理学杂志》2015年第2期291-296,共6页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81172710);深圳市科技研发资金基础研究计划(JC201105180762A);深圳市科技计划项目(201202097);广东省医学科研基金(A2014644)~~

摘　　要：目的探讨双酚A(BPA)对胚胎干细胞(ESC)分化潜能的影响,为合理评价BPA的安全性提供实验基础。方法制备及培养的小鼠胚胎成纤维细胞(MEF)和小鼠ESC,用BPA 0.1,1,10,100和1000μmol·L-1持续培养8 d,CCK-8法检测MEF和ESC细胞存活率并计算IC50;通过实时荧光定量PCR检测ESC中α肌球蛋白重链m RNA表达并计算其半数最大分化抑制浓度(ID50)。悬滴悬浮法培养的拟胚体,用BPA 0.001,0.01,0.1和1μmol·L-1持续培养10 d,实时荧光定量PCR检测拟胚体各胚层标志基因表达的变化。结果 BPA对小鼠ESC的IC50为5.22×10-4mol·L-1,对MEF的IC50为6.25×10-4mol·L-1,对小鼠ESC体外心肌细胞定向分化的ID50为7.0×10-7mol·L-1。BPA 0.001和0.01μmol·L-1可以上调拟胚体的中胚层标志基因胎肝激酶1和球蛋白转录因子1的表达。结论 BPA属于强胚胎毒性化合物。低浓度BPA对小鼠ESC分化为中胚层细胞有促进分化的作用。OBJECTIVE To explore the effect of bisphenol A （BPA） on the differentiation potential of embryonic stem cells, and provide an experimental basis for evaluation of safety of BPA. METHODS Mouse embryonic fibroblasts （MEFs） and embryonic stem cells （ESCs） were treated with BPA 0.1, 1, 10, 100 and 1000 μmol.L-1 for 8 d respectively. The viability of MEFs and ESCs was measured by CCK-8 and lC50 was calculated. The mRNA expression of α-myosin heavy chain in ESCs was tested by RT-PCR to determine lD50 . The embryonic body cultured by suspension method was treated with BPA 0.001, 0.01, 0.1 and 1 μmol.L-1 for 10 d respectively. The changes of marked genes in each blastoderm were detected by RT-PCR. RESULTS lC50 of BPA to mouse ESCs was 5.22×10-4 mol.L-1 , and to MEFs was 6. 25 × 10-4 mol.L-1 . lD50 of BPA to mouse ESCs differentiating to cardiomyocytes was 7.0×10-7 mol.L-1 . BPA 0.001 and 0.01 μmol.L-1 upregulated the expression of the marked genes of mesoderm, fetal liver kinase-1 and globin transcription factor 1. CONCLUSION BPA is a strong embry-otoxic compound. BPA of low concentration can promote the differentiation of mouse ESCs to mesoderm.

关 键 词：双酚A 胚胎干细胞 药物毒性 

分 类 号：R114[医药卫生—卫生毒理学]