应用胚胎干细胞实验模型评价氢醌的胚胎毒性  

Embryonic stem cell test for the study of the embryotoxicity of hydroquinone

作　　者：李付贵[1] 程伟民[1] 季明芳[1] 陈京[1] 

机构地区：[1]中山大学附属中山医院肿瘤研究所,广东中山528403

出　　处：《国际检验医学杂志》2015年第13期1807-1809,1812,共4页International Journal of Laboratory Medicine

基　　金：国家自然科学基金青年基金项目(81202604);中国博士后科学基金项目(2014M562244)

摘　　要：目的建立胚胎干细胞实验(EST)模型,应用胚胎干细胞实验模型初步评价氢醌的胚胎毒性。方法体外培养小鼠成纤维细胞3T3和小鼠胚胎干细胞E14TG2a(ES-E14TG2a),四甲基偶氮唑盐(MTT)法检测阳性对照5-氟尿嘧啶、阴性对照青霉素G和受试物氢醌对3T3细胞和ES-E14TG2a细胞的细胞毒性,计算3种化合物对ES-E14TG2a细胞和3T3细胞的半数生长抑制浓度(IC50)值IC50ES、IC503T3;采用悬滴-悬浮-贴壁法体外诱导胚胎干细胞向心肌细胞分化,根据浓度-反应曲线计算3种化合物对ES-E14TG2a细胞的半数分化抑制浓度(ID50)值ID50ES。利用胚胎毒性预测模型预测氢醌的胚胎毒性。结果氢醌对3T3细胞和E14TG2a细胞的增殖均有抑制作用,其IC503T3和IC50 ES分别为(5.97±0.48)、(2.57±0.10)μg/mL。氢醌对ESE14TG2a细胞的分化也有抑制作用,其ID50ES为(3.77±0.31)μg/mL。根据EST预测模型计算得出氢醌具有强胚胎毒性。结论氢醌具有强胚胎毒性。Objective To establish a model of embryonic stem cell test（EST）and utilize this model to evaluate the embryotoxici-ty of hydroquinone.Methods Mouse 3T3 fibroblasts and mouse embryonic stem（ES）cells（ES-E14TG2a）were cultured in vitro, and methyl thiazolyl tetrazolium（MTT）test was performed to detect the cytotoxicity of 3T3 cells and ES-E14TG2a cells induced by the positive control（5-fluorouracil）,negative control（penicillin G）and tested compound（hydroquinone）.The concentrations of the test compounds that inhibited 50% viability of ES-E14TG2a cells（IC50 ES）and 3T3 fibroblasts （IC50 3T3）were calculated.The hanging-suspension-adherent culture systems were used to induce embryonic stem cells into cardiomyocytes,and the concentrations of test compounds that caused 50% inhibition of differentiation of ES-E14TG2a cells into cardiomyocytes （ID50 ES）was calculated. The embryotoxic potential of hydroquinone was classified by prediction model of the embryonic stem cell test.Results The prolif-eration of ES-E14TG2a and 3T3 cells were inhibited by hydroquinone,of which the IC50 3T3 and IC50 ES values were （5.97±0.48） and （2.57±0.10）μg/mL respectively.The differentiation of ES-E14TG2a cells were also inhibited by hydroquinone,of which the ID50 ES was （3.77±0.31）μg/mL.Hydroquinone was evaluated as a strong embryotoxicity chemical by prediction model of EST. Conclusion Hydroquinone exhibits a strong embryotoxicity.

关 键 词：胚胎干细胞 氢醌 胚胎毒性 

分 类 号：R114[医药卫生—卫生毒理学]