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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 investigate the effects of DBT on the proliferation and differentiation of neural stem cells in rats during pregnancy. Methods Forty pregnant SD rats were randomly divided into experimental group and control group with three different doses of DBP [25,75,225 mg / (kg · d)]. The pregnant rats in each group were given different doses of DBP or corn oil solvent Gavage until production. Neonatal rats were used to culture neural stem cells. The colony formation rate and the size of the sphere were detected. The differentiated neural stem cells were differentiated and cultured. NeuN and GFAP immunofluorescence was used to detect the differentiation of neural stem cells into neurons and star Astrocytes ratio differences and morphological changes. Results The DBP induced progenitor rat neural stem cells proliferative capacity decreased during pregnancy. Compared with the control group, the clonality rate of NSCs in neonatal rats exposed to medium and high doses was significantly lower than that of the control group [(40.53 ± 4.65)% in the experimental control group and (30.96 ± 3.80) %, (15.35 ± 5.29)%, (6.58 ± 1.43)%, P <0.01]. The diameters of the cloned spheres in the high-dose treatment group were also significantly smaller than those in the control group. DBP high-dose group of neural stem cells differentiate into astrocytes significantly increased [high-dose group (36.8 ± 3.5)%, while the control group was (30.8 ± 2.4)%, P <0.01]. Differentiated neurons and glial cells morphology with the concentration increased, the morphological changes significantly. Astroglial cells gradually become shorter, thicker, high-dose exposure is obvious. Conclusion DBP during pregnancy may affect the proliferation and differentiation of neural stem cells in the offspring and affect the development of the nervous system.