环境雌激素EE2早期慢暴露对雄性大鼠成年后神经发育与空间学习行为的影响
The Effects of Chronic Exposure to EE2 in Early Life on Neurodevelopment and Spatial Learning Behavior in Adult Male Rats
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摘要: 环境雌激素17α-乙炔雌二醇(17alpha-ethynylestradiol, EE2)是一种常见的环境内分泌干扰物，主要经消化道被人和动物摄入，但其对中枢神经系统的影响尚不清楚。为探讨EE2早期慢暴露对Sprague-Dawley (SD)大鼠神经发育和成年后认知行为的影响及其机制，本研究选择5日龄雄性SD大鼠，随机分为3组：空白对照组(Control组)、EE2低剂量组(0.1 μg·kg-1)、EE2高剂量组(10 μg·kg-1)，每组8只，EE2持续暴露16周。观察大鼠体质量、脑质量变化。采用Morris水迷宫、旷场实验观察大鼠行为学表现。Nissl染色和免疫组织化学染色法观察各组大鼠海马CA1区(CA1)、海马CA3区(CA3)、齿状回(DG)的细胞形态和脑源性神经营养因子(BDNF)的表达，Western Blot检测海马、大脑皮层的IL-1β、IL-10蛋白水平，并测定皮层和海马丙二醛(MDA)含量。结果显示，EE2暴露组大鼠体质量增长速度较对照组减缓，脑质量减轻。行为学发现，EE2组逃逸潜伏期延长，目标象限停留时间、跨台次数减少，中央区运动距离和中央区停留时间减少，并呈剂量依赖性变化。EE2暴露组海马各区神经元排列紊乱、形态不规则，DG区神经元前体细胞数量减少。进一步发现，EE2暴露组大鼠大脑皮层、海马组织的MDA含量均升高，IL-1β表达量增加，IL-10表达量下降。低剂量组海马DG区BDNF表达减少，高剂量组CA1、CA3、DG区BDNF表达均减少。以上结果表明，生命早期0.1 μg·kg-1 EE2慢暴露可引起雄性大鼠成年后空间学习记忆行为障碍，与海马结构紊乱、BDNF水平降低和脑组织炎症反应有关。Abstract: 17alpha-ethynylestradiol (EE2), a prevalent environmental endocrine disruptor primarily ingested orally, leads to its accumulation in the body. This study aimed to investigate the effects of neonatal exposure to EE2 on neurodevelopment and cognitive behavior in adulthood, as well as the underlying mechanism. Male Sprague-Dawley rats were exposed to low-dose EE2 (0.1 μg·kg-1), high-dose EE2 (10 μg·kg-1), or control for 16 weeks. The alterations in body weight and whole brain weight were determined, while neurobehavioral assessment was conducted using Morris water maze and Open field test. Nissl staining and immunohistochemistry were employed to observe morphology and expression of brain-derived neurotrophic factor (BDNF) in hippocampal regions CA1, CA3, and DG; Western Blot and biochemical assay measured levels of IL-1β, IL-10 protein, and malondialdehyde (MDA) in hippocampal/cortical tissues. The results showed that rats exposed to EE2 had lower weight growth and brain weight. Furthermore, the neurobehavioral performance indicated longer escape latency, reduced residence time in target quadrant and cross-platform times, also decreased movement distance in the central area, and less central residence time. These variations exhibited a dose-response relationship. Disorganized hippocampal neurons, irregular cell shapes, and a decreased number of neuronal precursor cells were noticed in the DG area of the hippocampus compared with controls while elevated levels of IL-1β and decreased levels of IL-10 as well as increased MDA levels detected both cerebral cortex/hippocampus among rats exposed to EE2 along with decreased BDNF expression in DG region for the low dose group and throughout entire region for high dose group. These findings imply that chronic exposure to EE2 at a dose of 0.1 μg·kg-1 during early-life in male rats can result in spatial learning deficits in adulthood, which might be associated with disorders in hippocampal formation, lower BDNF expression, and inflammation in both the cerebral cortex and hippocampus.
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Key words:
- 17α-ethynylestradiol /
- rat /
- early chronic exposure /
- hippocampus /
- learning and memory behavior
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