胚胎干细胞试验模型用于白藜芦醇胚胎发育毒性的评价

The evaluation of resveratol's embryotoxicity by embryonic stem cell test model

目的依据胚胎干细胞(Embryonic Stem Cell,ESC)具有向心肌细胞分化的能力,建立体外分化发育毒性评价模型,并应用其评价受试物白藜芦醇胚胎发育毒性。方法悬滴转悬浮法培养D3系胚胎干细胞(ESC-D3),观察受试物对ESC-D3分化能力影响,结合MTT法判断受试物对ESC-D3及小鼠胚胎成纤维细胞(3T3)的细胞毒性结果,预测受试物的发育毒性。用已知无胚胎发育毒性化合物青霉素(Penicillin,PN),强胚胎发育毒性化合物氟尿嘧啶(5-Fluorouracil,5-FU)等对模型进行有效性验证,并将经过验证的EST模型用于受试物白藜芦醇的发育毒性评价。结果利用建立的EST模型对5-FU和PN的发育毒性进行评价,结果为5-FU为强胚胎发育毒性,PN为无胚胎发育毒性,与文献报道一致。应用EST模型评价得出白藜芦醇对3T3细胞活力的半数抑制浓度IC503T3为27.93μg/ml,对D3细胞活力的半数抑制浓度IC50D3为28.02μg/ml,对D3细胞的半数分化抑制浓度ID50D3为158.97μg/ml,经EST模型判断公式计算得出该化合物无发育毒性化合物。结论建立的EST模型的有效性验证结果与ECVAM的结论一致,可用于胚胎发育毒性的筛选和评价;经EST模型评价,白藜芦醇为无胚胎发育毒性。

Objective According the potency and trend of ESC differentiating to cardiomyocyte, the model of EST had been built and the embryonic toxicity of resveratrol was evaluated by EST model. Methods On the basis of endpoint of inhibition of differentiation of ESC into cardiac myoblasts and the endpoints of inhibition of cell growth in ESC and 3T3 cells, test chemical could be classified as non, weak or strong embryotoxic by a predicted model. After the validation of the EST model was tested by the non embryotoxie chemical, Penicillin G （PN）, and strong embryotoxic chemical, Fluorouracil （5-FU） , the embryonic toxicity of resveratrol was evaluated by the EST model. Results PN was discriminated as non embryotoxicity and and 5-FU was discriminated as strong Embryotoxieity, the results were coincide with the predication by ECVAM. The endpoints of resveratrol were IC5o3T3 （27.93 μg/ml） ,ICsoD3 （28.02 μg/ml） and IDsoD3 （158.97 μg/ml） and resveratrol was evaluated as a non embryotoxicity chemical by EST. Conclusion The validation of EST model established by us was high and resveratrol was evaluated as a non embryotoxicity chemical by EST.