大鼠孕期染邻苯二甲酸二丁酯对子代神经干细胞增殖分化的影响  

Effect of maternal exposure to dibutyl phthalate on proliferation and differentiation of neural stem cells from neonatal offspring rats

作　　者：罗芬兰[1] 蔡文琴[1] 张吉强[1] 

机构地区：[1]第三军医大学基础医学部神经生物学教研室,重庆市神经科学研究所,重庆400038

出　　处：《第三军医大学学报》2010年第11期1188-1191,共4页Journal of Third Military Medical University

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、225mg/(kg·d)]的实验组和对照组,各组孕鼠自E0起分别给予不同剂量DBP或玉米油溶剂灌胃直至生产。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干预后的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)%,P<0.01]。高剂量处理组克隆球的直径也明显小于对照组。DBP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)%,P<0.01]。分化后的神经元和神经胶质细胞形态随着染毒浓度增大,形态变化明显。星形胶质细胞的细胞突起逐渐变短,变粗,以高剂量染毒后为明显。结论大鼠孕期染DBP会影响子代神经干细胞的增殖与分化,从而影响神经系统的发育。Objective To explore the effects of maternal exposure to dibutyl phthalate （ DBP） ,an environmental estrogen on the proliferation and differentiation of neural stem cells （ NSCs） in the neural tube of neonatal rats. Methods A total of 40 male and 40 female SD rates at age of 4 to 5 months,were matched,and the morning when vaginal plugs were found was designated as E0. Then the 40 pregnant rats were randomly divided into 4 groups,3 DBP exposure groups （ 25,75,and 225 mg/kg） and control group. DBP at corresponding doses were dissolved in corn oil,and administrated by intragastrically injection once per day from E0 till delivery,while those of control group were given corn oil. Brain tissue of newborn rats was collected for primary culture of NSCs. Then the cell colony formation was counted to detect the NSCs proliferation. Then 10% fetal bovine serum （ FBS） was added into the culture medium to induce the NSCS to differentiate. Neu-N and GFAP immunofluorescence stainings were used to detect neurons and astrocytes,and their morphological changes were observed. Results The proliferation of NSCs was decreased significantly after DBP administration. Compared with the control group （ 40. 53 ± 4. 65） % ,the colony formation rate of middle-and high-dose DBP exposure groups was significantly reduced and in a dose-dependent manner （ 30. 96 ± 3. 80） % ,（ 15. 35 ±5. 29） %,（ 6. 58 ±1. 43） %,P 0. 01. The diameter of the formed colony in high-dose DBP exposure group was significantly smaller than in control group. DBP induced more NSCs to differentiate to astrocytes in high-dose group （ 36. 8 ± 3. 5） % than in control group （ 30. 8 ± 2. 4） % ,P 0. 01Morphological changes were more obvious in the differentiated neurons and astrocytes with the increasing dose of DBP. DBP also caused obvious gradually shortened and thickened processes,especially in the high dose group. Conclusion The results suggest that DBP exert obvious effect on the proliferation and differentiations of NSCs,and thus

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 

分 类 号：R715.3[医药卫生—妇产科学] R322.8[医药卫生—临床医学]