双酚A对小鼠胚胎干细胞分化潜能的影响
发布时间：2018-09-02 05:37
【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。
[Abstract]:Objective to investigate the effect of bisphenol A (BPA) (BPA) on the differentiation potential of embryonic stem cells (ESC), and to provide an experimental basis for evaluating the safety of BPA. Methods (MEF) of mouse embryonic fibroblasts and ESC, of mice were prepared and cultured. The survival rate of MEF and ESC cells was detected by BPA 0.1 渭 mol L-1 and 1000 渭 mol L-1 CCK-8 method for 8 days. IC50; was used to detect 伪 -myosin heavy chain m RNA in ESC by real-time fluorescence quantitative PCR. The maximum inhibitory concentration of differentiation (ID50) was calculated. The embryoid bodies cultured with suspension method were incubated with BPA 0.001 0. 01 and 1 渭 mol L-1 for 10 days. The expression of marker genes in each embryo layer was detected by real time fluorescence quantitative PCR. Results the IC50 of BPA on ESC was 5.22 脳 10-4mol L-1, IC50 on MEF was 6.25 脳 10-4mol L-1. ID50 of directed differentiation of mouse ESC in vitro was 7.0 脳 10-7mol L-1.BPA 0.001 and 0.01 渭 mol L-1. The expression of fetal liver kinase 1 and globulin transcription factor 1 was up-regulated by fetal liver kinase 1 and globulin transcription factor 1. Conclusion BPA is a strong embryotoxic compound. Low concentration of BPA can promote the differentiation of mouse ESC into mesodermal cells.
【作者单位】： 福建医科大学公共卫生学院环境与健康重点实验室;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;深圳市慢性病防治中心;
【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)~~
【分类号】：R114
[Abstract]:Objective to investigate the effect of bisphenol A (BPA) (BPA) on the differentiation potential of embryonic stem cells (ESC), and to provide an experimental basis for evaluating the safety of BPA. Methods (MEF) of mouse embryonic fibroblasts and ESC, of mice were prepared and cultured. The survival rate of MEF and ESC cells was detected by BPA 0.1 渭 mol L-1 and 1000 渭 mol L-1 CCK-8 method for 8 days. IC50; was used to detect 伪 -myosin heavy chain m RNA in ESC by real-time fluorescence quantitative PCR. The maximum inhibitory concentration of differentiation (ID50) was calculated. The embryoid bodies cultured with suspension method were incubated with BPA 0.001 0. 01 and 1 渭 mol L-1 for 10 days. The expression of marker genes in each embryo layer was detected by real time fluorescence quantitative PCR. Results the IC50 of BPA on ESC was 5.22 脳 10-4mol L-1, IC50 on MEF was 6.25 脳 10-4mol L-1. ID50 of directed differentiation of mouse ESC in vitro was 7.0 脳 10-7mol L-1.BPA 0.001 and 0.01 渭 mol L-1. The expression of fetal liver kinase 1 and globulin transcription factor 1 was up-regulated by fetal liver kinase 1 and globulin transcription factor 1. Conclusion BPA is a strong embryotoxic compound. Low concentration of BPA can promote the differentiation of mouse ESC into mesodermal cells.
【作者单位】： 福建医科大学公共卫生学院环境与健康重点实验室;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;深圳市慢性病防治中心;
【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)~~
【分类号】：R114
【参考文献】
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