碱性橙Ⅱ对大鼠胚胎肢芽细胞增殖和分化的影响

Effects of basic orange Ⅱ on proliferation and differentiation of limb bud cells in rat embryos

目的探讨碱性橙Ⅱ对13日龄SD大鼠胚胎肢芽细胞增殖和分化的影响。方法分离13日龄SD大鼠胚胎肢芽细胞进行微团培养,不同浓度的碱性橙Ⅱ(0.0、12.5、25.0、50.0、100.0、200.0和400.0 mg/L)作用于肢芽细胞,采用CCK8试剂盒检测碱性橙Ⅱ对细胞增殖的影响,采用阿利新蓝8GX染色方法检测碱性橙Ⅱ对肢芽细胞分化的影响。结果随着碱性橙Ⅱ染毒浓度的增加,胚胎肢芽细胞分化为软骨细胞集落数逐渐减少,细胞增殖率下降,均具有较明显的剂量-效应关系(r细胞毒性=0.958,r细胞分化=0.946);碱性橙Ⅱ对肢芽细胞的半数增殖抑制浓度(p ID50)和半数分化抑制浓度(d ID50)分别为240.6和69.3 mg/L,对细胞分化的抑制作用大于对增殖的抑制作用。结论碱性橙Ⅱ对胚胎肢芽细胞的增殖和分化有抑制作用,可能对大鼠胚胎具有潜在的发育毒性。

Objective To explore the effects of basic orangeⅡ on proliferation and differentiation of limb bud cells. Methods Limb bud cell were separated from SD rat embryo at 13-day gestational age, limb bud cell were exposed to basic orange Ⅱ at concentrations of 0.0,12.5,25.0,50.0,100.0,200, 0 and 400.0 mg/L in the culture medium. The effect of basic orange II on limb bud cell proliferation was detected by Cell Counting Kit-8, the effect of basic orange II on limb bud cell differentiation was assessed by Aleian Blue 8GX. Results With the increasing of basic orange ]I concentration, the proliferation and differentiation of embryo limb bud cells were poorer and poorer in vitro, and there was the dose-effect relationship. The pIDs0 and dIDso of basic orange Ⅱ on limb bud cells were 240. 6 mg/L and 69. 3 mg/L respectively. The inhibition of basic orange Ⅱ on cell differentiation might exceed that on cell proliferation. Conclusions Basic orange Ⅱ could inhibit proliferation and differentiation of embryo limb bud cells. It might be a potential developmental toxic substance in rat embryo.