Analysis for the mechanisms affecting neural of development by endocrine disrupting chemicals using non-human primates

考虑到在进行内分泌干扰化学品对人类的风险评估时的物种差异，实验中使用了密切相关的非人类灵长类动物。根据前人的研究结果，采用双酚A作为雌激素制剂，采用甲巯咪唑替代具有抑制甲状腺激素作用的多氯联苯或TCDD。双酚A处理组雄性和雌性后代分别进行了脑两性二态核的蛋白质组分析。暴露组男性表现为女性模式，暴露组女性表现为中性模式。针对与对照组存在显著差异的肽点，研究暴露组的性别差异。用免疫组织化学方法检测了暴露组和对照组中覆盖靶点的几种蛋白质的性别差异，然而，从形态学上很难发现对照组性别差异或暴露效应之间的差异。今后应进行定量分析，包括激素受体。甲巯咪唑给药实验发现，严重影响胎儿甲状腺激素的产生和大脑发育的非人灵长类动物。对甲巯咪唑的作用机理进行了实验研究。在妊娠的不同阶段（妊娠早期、妊娠中期、妊娠晚期和妊娠末期）给母鼠施甲巯咪唑，并对个体获得的子代胎儿脑进行精确分析。妊娠末期（妊娠130 ~ 150天）治疗影响最大。因此，我们进行了新的实验，在此基础上增加了4个个体，以获得可重复的结果。Western blot法定量分析候选蛋白表达变化。暴露组可观察到甲状腺肿大和功能障碍，重现性明显，但在神经元发育方面，蛋白表达水平的个体差异较大。暴露组有大神经细胞突起发育迟缓的趋势，但差异无统计学意义。甲状腺激素浓度与个体甲状腺增大和神经发育迟缓之间的相关性并没有明确的关系。与其他神经细胞相比，小脑浦肯野细胞和大脑皮层锥体细胞受到暴露的强烈影响。今后对这些细胞进行定量分析是必要的。少

Consideration of species differences in conducting risk assessments of endocrine disrupting chemicals to humans, closely related non-human primates were used in experiments. Bisphenol A was used as an estrogenic agents from the results of previous studies, and methimazole was conducted as an alternative of the PCB or TCDD which have the effect of inhibiting thyroid hormones. Bisphenol A treated groups consisting of male and female offspring were done in proteome analysis for sexually dimorphic nucleus of the brain. Exposure group of male exhibited female patterns, and exposed females tended to show neutral patterns. Peptide spots showing significant differences from the control group were targeted, and sex differences in the exposed group were studied. Several proteins covering the targeted spots were examined by immunohistochemical methos in both exposure and control groups for sex differences, however, to find the difference between the control sex difference or exposure effects were … More difficult morphologically. The quantitative analysis should be carried, including hormone receptors in the future. Methimazole administration experiments were found to affect severely both fetal thyroid hormone production and brain development in non-human primates. The experiments to elucidate the mechanisms of methimazole were proceeded. During different stages of pregnancy (early pregnancy, mid, late, and end stages) mothers were administered methimazole and the offspring fetal brain obtained from individuals were analyzed precisely. Terminal phase (doses from the 130 to 150 days in gestation) treatments influenced the strongest effect. Thus, new experiment was proceeded to add four more individuals to obtain reproducible results based on this. Quantitative analysis of changes in the expression of candidate proteins by Western blot analysis. Thyroid enlargement and dysfunction were observed in the exposed group with a clear reproducibility, however, as for neuronal development, individual differences in protein expression level were strong. A tendency of retardation in large neural cell projections in the exposed group was observed but there was no significant difference. A correlation between thyroid hormone concentrations with enlarging thyroid glands and neural developmental retardations in individuals did not reveal a clear relationship. Purkinje cells in the cerebellar and pyramidal cells in the cerebral cortex showed a strong impact by exposure compared to other neural cells. An experiment targeted to these cells was considered necessary by quantitative analysis of individual in the future. Less