CAJ | 학술논문

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 lC50；通过实时荧光定量 PCR检测 ESC 中α肌球蛋白重链 mRNA 表达并计算其半数最大分化抑制浓度(lD50)。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 lC50为5.22×10-4 mol??L-1,对 MEF 的 lC50为6.25×10-4 mol??L-1,对小鼠 ESC 体外心肌细胞定向分化的 lD50为7.0×10-7 mol??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。
목적：탐토쌍분 A(BPA)대배태간세포(ESC)분화잠능적영향,위합리평개 BPA 적안전성제공실험기출。방법제비급배양적소서배태성섬유세포(MEF)화소서 ESC,용 BPA 0.1,1,10,100화1000μmol??L-1지속배양8 d,CCK-8법검측 MEF 화 ESC 세포존활솔병계산 lC50；통과실시형광정량 PCR검측 ESC 중α기구단백중련 mRNA 표체병계산기반수최대분화억제농도(lD50)。현적현부법배양적의배체,용 BPA 0.001,0.01,0.1화1μmol??L-1지속배양10 d,실시형광정량 PCR 검측의배체각배층표지기인표체적변화。결과 BPA 대소서 ESC 적 lC50위5.22×10-4 mol??L-1,대 MEF 적 lC50위6.25×10-4 mol??L-1,대소서 ESC 체외심기세포정향분화적 lD50위7.0×10-7 mol??L-1。 BPA 0.001화0.01μmol??L-1가이상조의배체적중배층표지기인태간격매1화구단백전록인자1적표체。결론 BPA 속우강배태독성화합물。저농도 BPA 대소서 ESC 분화위중배층세포유촉진분화적작용。
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.