中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2019, Vol. 14 ›› Issue (5): 834-840.doi: 10.4103/1673-5374.249231

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

出生后给予calpeptin可导致乳鼠海马神经发育缺陷

  

  • 出版日期:2019-05-15 发布日期:2019-05-15
  • 基金资助:

    国家自然科学基金项目(81601181, 81673716)；安徽省自然科学基金项目(1808085J15)；安徽省自然科学研究项目KJ2016A417)

Postnatal calpeptin treatment causes hippocampal neurodevelopmental defects in neonatal rats

Zhu-Jin Song 1 , San-Juan Yang 1 , Lan Han 2 , Bin Wang 1 , Guoqi Zhu 1   

  1. 1 Key Laboratory of Xin’an Medicine, Ministry of Education, Anhui University of Chinese Medicine, Hefei, Anhui Province, China
    2 School of Pharmacy, Anhui University of Chinese Medicine, Hefei, Anhui Province, China
  • Online:2019-05-15 Published:2019-05-15
  • Contact: Guoqi Zhu, PhD, guoqizhu@gmail.com.
  • Supported by:

    This study was supported by the National Natural Science Foundation of China, No. 81601181, 81673716 (to GZ); the Natural Science Foundation of Anhui Province of China, No. 1808085J15 (to GZ); and the Natural Science Research Project of Anhui Province of China, No. KJ2016A417 (to GZ).

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摘要：

作者既往研究结果表明，早期使用钙蛋白酶抑制剂会降低多个脑区中钙蛋白酶的活性，而出生后应用calpeptin可能会导致小脑运动功能障碍。然而，产后使用calpeptin是否影响海马相关行为改变及其机制尚不清楚。为此，实验选用购自安徽医科大学动物中心的SD大鼠。(1)成年阶段的实验：SD大鼠在实验第7-14天时每天腹腔注射1次calpeptin（2mg/kg）；以Morris水迷宫测试空间学习记忆能力，以旷场试验测试焦虑样活动，以鬼笔环肽染色测量海马中突触形态，以免疫组化染色检测海马CA1区NeuN阳性细胞的数量，以DiI标记树突棘。结果发现给予calpeptin，会损害空间记忆，并引起成年期焦虑样行为，同时减少了海马中顶端树突棘和肌动蛋白聚合的数量和面积，但未影响海马CA1区中NeuN阳性细胞的数量；(2)出生后阶段的实验：将乳鼠分别在出生后第7和8天时腹腔注射calpeptin（2 mg/kg），以Western blot分析乳鼠海马中Akt，Erk，p-Akt，p-Erk1/2，Erk1/2，SCOP，PTEN，mTor，p-mTor，CREB、p-CREB的表达水平。发现大鼠海马中SCOP水平增加，Akt磷酸化水平降低，mTor和CREB磷酸化程度下降。表明出生后给予calpeptin可能通过抑制Akt/mTor信号通路，影响乳鼠突触发育，继而损害海马神经发育，提示出生后应用calpeptin是神经发育缺陷的风险因素。

orcid: 0000-0001-7432-6079 (Guoqi Zhu)

关键词: 产后, calpeptin, 学习记忆, 海马, 树突棘发育, SCOP, AKT, mTOR, 神经再生

Abstract:

Our previous studies showed that the early use of calpain inhibitors reduces calpain activity in multiple brain regions, and that postnatal treatment with calpeptin may lead to cerebellar motor dysfunction. However, it remains unclear whether postnatal calpeptin application affects hippocampus-related behaviors. In this study, Sprague-Dawley rats were purchased from the Animal Center of Anhui Medical University of China. For the experiments in the adult stage, rats were intraperitoneally injected with calpeptin, 2 mg/kg, once a day, on postnatal days 7–14. Then on postnatal day 60, the Morris water maze test was used to evaluate spatial learning and memory abilities. The open field test was carried out to assess anxiety-like activities. Phalloidin staining was performed to observe synaptic morphology in the hippocampus. Immunohistochemistry was used to count the number of NeuN-positive cells in the hippocampal CA1 region. DiI was applied to label dendritic spines. Calpeptin administration impaired spatial memory, caused anxiety-like behavior in adulthood, reduced the number and area of apical dendritic spines, and decreased actin polymerization in the hippocampus, but did not affect the number of NeuN-positive cells in the hippocampal CA1 region. For the neonatal experiments, neonatal rats were intraperitoneally injected with calpeptin, 2 mg/kg, on postnatal days 7 and 8. Western blot assay was performed to analyze the protein levels of Akt, Erk, p-Akt, p-Erk1/2, Erk1/2, SCOP, PTEN, mTOR, p-mTOR, CREB and p-CREB in the hippocampus. SCOP expression was increased, and the phosphor¬ylation levels of Akt, mTOR and CREB were reduced in the hippocampus. These findings show that calpeptin administration after birth affects synaptic development in neonatal rats by inhibiting the Akt/mTOR signaling pathway, thereby perturbing hippocampal function. Therefore, calpeptin administration after birth is a risk factor for neurodevelopmental defects.

Key words: nerve regeneration, postnatal, calpeptin, learning and memory, hippocampus, spine development, SCOP, AKT, mTOR, neural regeneration