摘要
五氧化二钒(V2O5)是一种常见的工业毒物和环境污染物。为了进一步验证钒的致畸性,并为揭示其发育毒性的作用机理提供依据,本文在大鼠致畸试验的基础上,应用细胞微团培养方法观察了钒对大鼠胚胎肢芽细胞在体内外增殖和分化的影响。体外实验:从妊娠d13的孕鼠中分离出胚胎,取前肢芽,制成细胞悬液(2x107cel/mL),取10μL置于培养板上成一微团,于高湿、37℃的CO2孵箱中预培养2h,然后缓慢加入不同剂量的V2O5溶液(勿冲散微团,对照加培养基),培养5d。体内实验:取鼠妊娠d12时,经i.p,以不同剂量的V2O5处理,16h后处死、取胚胎肢芽细胞培养。然后,固定细胞,分别采用苏木精染色和阿利新兰染色,前者经甲醇脱色后以酶标光密度值反映细胞增殖情况;后者由镜检每一微团中天蓝色的分化集落数判断对细胞分化的抑制作用。体外试验结果显示,钒剂量在5.0μmol/L以上时细胞集落数明显下降,光密度则在7.5μmol/L以上时明显下降,均呈明显的剂量反应关系(Table1),说明钒对肢芽细胞增殖和分化都有抑制作用,但细胞增殖和分化的两条抑制曲线明显不同(Fig.1),其IC50分别为13.14和4.77μmol/L。按F?
The micromass culture was used to determine the effects of vanadium pentoxide (V2O5 ) on the proliferation and differentiation of limb bud cells of rat. In the in vitro test, the results showed that V2O5 had obvious inhibiting effects on both proliferation and differentiation of limb bud cells with a dosedependent response, its proliferating and differentiating IC50 being 13.64 and 4.77μmol/L, respectively. In the in vivo/in vitro test, the results showed that V2O5 had no obvious effect on cell proliferation but had obvious inhibiting effect on cell differentiation. These results indicated that V2O5 might have a specific inhibiting effect on the differentiation of limb bud cells.
关键词
五氧化二钒
大鼠
胚胎
肢芽细胞增殖
分化
致畸性
micromass culture, limb bud cells, developmental toxicity, V2O5, rat.