孕大鼠染邻苯二甲酸二丁酯对子代神经干细胞增殖分化影响.doc

孕大鼠染邻苯二甲酸二丁酯对子代神经干细胞增殖分化影响.doc

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孕大鼠染邻苯二甲酸二丁酯对子代神经干细胞增殖分化的影响 罗芬兰,蔡文琴,张吉强(第三军医大学神经生物学教研室,重庆市神经科学研究所,重庆400038) [摘要] 目的 探讨大鼠孕期染环境雌激素邻苯二甲酸二丁酯(dibutyl phthalate,DBP)对子代神经干细胞增殖和分化的影响。方法 40只SD孕鼠随机分为3个不同DBP剂量[25、75、225 mg/(kg·d)]的实验组和溶剂对照组,各组孕鼠自E0起分别给予不同剂量DBP或溶剂灌胃直至生产。取新生大鼠进行神经干细胞培养,检测计数细胞克隆形成率、克隆球直径大小;分离的神经干细胞进行分化培养,以Neu-N和GFAP免疫荧光检测DBP神经干细胞分化为神经元与星形胶质细胞的比例差别与形态变化。结果 大鼠孕期染DBP致子代神经干细胞增殖能力降低。与对照组相比,中、高剂量染毒组新生鼠神经干细胞克隆率明显低于对照组[实验对照组克隆率为(40.53+4.65)%;药物处理组克隆率分别为(30.96+3.80)%、(15.35+5.29)%、(6.58+1.43)%,P0.01)]。高剂量处理组克隆球的直径也明显小于实验对照组(P0.05)。DBP处理使高剂量组神经干细胞分化为星形胶质细胞比例显著增加[高剂量组为(36.8±3.5)%,而对照组为(30.8±2.4)%,P0.01 ]。分化后的神经元和神经胶质细胞形态随着毒物剂量增大,形态变化明显。神经元与星形胶质细胞,细胞突起逐渐变短,变粗,以高剂量染毒后为明显。结论 DBP会影响大鼠神经干细胞的增殖与分化,从而影响神经系统的发育。 [关键词] 环境雌激素;邻苯二甲酸二丁酯;神经干细胞;神经干细胞增殖与分化 Effect of pregnant rats exposure to dibutyl phthalate on the proliferation and differentiation of neural stem cells in neonatal rats Luo Fenlan,Cai Wenqin,Zhang Jiqiang(Department of Neurobiology, Chongqing Neuroscience Institute, Chongqing 400038) [Abstract] Objective This study was designed to explore the effects of dibutyl phthalate (DBP) on proliferation and differentiation of neural stem cells (NSCs) from neural tube of embryonic rats. Methods 40 male and 40 female SD rates of 4-5 moths age, the rates were matched then the vaginal plugs were examined every morning, the day when discovering vaginal plug was designated as E0. Pregnant rats were randomly divided into four groups: 3 DBP exposed groups (DBP dose: 25-,75-and 225-mg/kg/d, respectively; dissolved in corn oil, administrated by digestive tract from E0 to the newborn pups) and normal control group (pregnant rats were given corn oil). Brain tissue of newborn rats was collected and the NSCs were cultured, the cell colony formation was counted to detect the NSCs proliferation. Then, with 10% fetal bovine serum (FBS), the ratio of the differentiated neurons to glia cells and morphological changes were studied. Results The proliferation of NSCs decreased significantly after DBP administration. The differen

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