应用微团培养和胚胎干细胞试验模型评价二硝酰胺铵的发育毒性  

Evaluation of developmental toxicity of ammonium dinitramide by micromass culture and embryonic stem cells models

作　　者：高俊宏 王鸿 范小琳 刘志永 赵彬 高婷 Gao Junhong;Wang Hong;Fan Xiaolin;Liu Zhiyong;Zhao Bin;Gao Ting(Institute for Hygiene of Ordnance Industry,Xi'an 710065,China)

机构地区：[1]兵器工业卫生研究所,西安710065

出　　处：《中华劳动卫生职业病杂志》2021年第11期815-818,共4页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金："十三五"预研项目(YY50201)。

摘　　要：目的利用微团培养和胚胎干细胞试验评价二硝酰胺铵(ADN)潜在的胚胎发育毒性,并评估其是否为致畸物。方法于2018年9月,分离大鼠胚胎并收集肢芽细胞,使用不同浓度(0、312.50、625.00、1250.00、2500.00、5000.00、10000.00μg/ml)ADN进行染毒,计算细胞半数增殖抑制浓度和半数分化抑制浓度并评价ADN致畸作用。在胚胎干细胞试验中,检测不同浓度(0、39.06、78.13、156.25、312.50、625.00、1250.00、2500.00μg/ml)ADN对小鼠胚胎干细胞(mESCs)向心肌细胞分化的抑制作用,以及对mESCs和3T3细胞的细胞毒性,并评价其胚胎毒性。以已知的强胚胎毒性药物5-氟尿嘧啶和非胚胎毒性药物青霉素-G为试验材料,对模型的有效性进行验证,并采用验证性试验模型评价ADN的胚胎毒性。结果微团培养试验中,各个浓度ADN组大鼠胚胎肢芽细胞增殖抑制率和分化抑制率均高于对照组(P<0.05);ADN对肢芽细胞的半数增殖抑制浓度和半数分化抑制浓度分别为7480.32和4526.09μg/ml,判定ADN为非致畸物。胚胎干细胞试验中,各个浓度ADN组mESCs增殖抑制率高于对照组,156.25、312.50、625.00、1250.00、2500.00μg/ml ADN组3T3细胞增殖抑制率高于对照组(P<0.05)。ADN对mESCs的半数增殖抑制浓度和半数分化抑制浓度分别为1851.73、1796.39μg/ml,对3T3细胞的半数增殖抑制浓度为3334.35μg/ml,判定ADN为无胚胎毒性。结论经微团培养和胚胎干细胞试验模型评价,ADN无胚胎毒性,属于非致畸物。Objective To evaluated the potential developmental toxicity and teratogenicity of ammonium dinitroamide(ADN)by micromass test(MM Test)and embryonic stem cell test models.Methods In September 2018,rat embryos were isolated and limb bud cells were collected.The limb bud cells were treated with different concentrations of ADN(0,312.50,625.00,1250.00,2500.00,5000.00,10000.00μg/ml).Half proliferation inhibitory concentration and half differentiation inhibitory concentration were calculated and the teratogenic effects were evaluated according to the criteria.For the embryonic stem cell test,the effects of different concentrations of ADN(0,39.06,78.13,156.25,312.50,625.00,1250.00,2500.00μg/ml)on the differentiation of mouse embryonic stem cells(mESCs)into myocardial cells and the cytotoxicity of mESCs and 3T3 cells were detected.The embryonic toxicity was evaluated according to the criteria.In this study,both 5-fluorouracil(5-FU),a known strong embryonic toxic drug,and penicillin-G(P-G),a non-embryonic toxic drug,were used to verify the effectiveness of the model,and the validated test model was applied to evaluate the embryonic toxicity of ADN.Results In the MM Test,the inhibition rates of proliferation and differentiation of limb bud cells in ADN groups were higher than that in control group(P<0.05).And the half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on limb bud cells were 7480.32 and 4526.09μg/ml,respectively.ADN was determined to be non-teratogenic by standard.In the embryonic stem cell test,the inhibition rates of mESCs proliferation in ADN groups were higher than that in control group,and the inhibition rates of 3T3 cells in 156.25,312.50,625.00,1250.00,2500.00μg/ml ADN groups were higher than that in control group(P<0.05).The half proliferation inhibitory concentration and half differentiation inhibitory concentration of ADN on mESCs were 1851.73 and 1796.39μg/ml,respectively,and the half proliferation inhibitory concentration on 3T3 cells was 3334.35�

关 键 词：胚胎干细胞 二硝酰胺铵 微团培养 发育毒性 大鼠 

分 类 号：R992[医药卫生—毒理学]