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利用胚胎干细胞试验模型评价矮壮素的发育毒性

Keywords: 矮壮素,植物生长调节剂,发育毒性,胚胎干细胞试验模型

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Abstract:

?目的:利用胚胎干细胞试验模型评价矮壮素的发育毒性。方法:以dmso为阴性对照,使用不同剂量(7.8125、15.625、31.25、62.5、125、250、500、1000μg/ml)的矮壮素染毒小鼠胚胎干细胞d3(es-d3)和3t3成纤维细胞,观察矮壮素对es-d3细胞向心肌细胞分化的影响,以及对es-d3和3t3细胞的细胞毒性,在此基础上,根据发育毒性判别公式对矮壮素致畸性进行判定。提取es-d3分化抑制试验中的拟胚体总rna,进行real-timepcr,在基因水平检测矮壮素对es-d3向各胚层分化的影响。结果:1000μg/ml矮壮素染毒可明显促进体外培养es-d3、3t3细胞增殖;250μg/ml的矮壮素作用下,观察到具有心肌细胞收缩的拟胚体数仅占接种拟胚体总数的69.4%;1000μg/ml矮壮素能够影响es-d3向心肌细胞分化的基因标志物(nkx2.5,α-mhc)的表达。结论:无法利用判别公式对矮壮素进行发育毒性评价;矮壮素在不引起3t3、es-d3细胞毒性的水平下,影响es-d3向心肌细胞的分化,有产生胚胎毒性的可能。

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