【摘要】：目的探討雙酚A(BPA)對胚胎干細胞(ESC)分化潛能的影響,為合理評價BPA的安全性提供實驗基礎(chǔ)。方法制備及培養(yǎng)的小鼠胚胎成纖維細胞(MEF)和小鼠ESC,用BPA 0.1,1,10,100和1000μmol·L-1持續(xù)培養(yǎng)8 d,CCK-8法檢測MEF和ESC細胞存活率并計算IC50;通過實時熒光定量PCR檢測ESC中α肌球蛋白重鏈m RNA表達并計算其半數(shù)最大分化抑制濃度(ID50)。懸滴懸浮法培養(yǎng)的擬胚體,用BPA 0.001,0.01,0.1和1μmol·L-1持續(xù)培養(yǎng)10 d,實時熒光定量PCR檢測擬胚體各胚層標志基因表達的變化。結(jié)果 BPA對小鼠ESC的IC50為5.22×10-4mol·L-1,對MEF的IC50為6.25×10-4mol·L-1,對小鼠ESC體外心肌細胞定向分化的ID50為7.0×10-7mol·L-1。BPA 0.001和0.01μmol·L-1可以上調(diào)擬胚體的中胚層標志基因胎肝激酶1和球蛋白轉(zhuǎn)錄因子1的表達。結(jié)論 BPA屬于強胚胎毒性化合物。低濃度BPA對小鼠ESC分化為中胚層細胞有促進分化的作用。
[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 偽 -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.
【作者單位】： 福建醫(yī)科大學公共衛(wèi)生學院環(huán)境與健康重點實驗室;深圳市疾病預防控制中心現(xiàn)代毒理學重點實驗室深圳市衛(wèi)生毒理學重點實驗室;深圳市慢性病防治中心;
【基金】：國家自然科學基金(81172710) 深圳市科技研發(fā)資金基礎(chǔ)研究計劃(JC201105180762A) 深圳市科技計劃項目(201202097) 廣東省醫(yī)學科研基金(A2014644)~~
【分類號】：R114

【參考文獻】

相關(guān)期刊論文 前3條

1 逄兵,周袁芬,周天喜,任道風,金泰^，

本文編號：2218417