中国神经再生研究(英文版) ›› 2017, Vol. 12 ›› Issue (10): 1648-1654.doi: 10.4103/1673-5374.217345

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

丙烯酰胺对断乳期幼鼠海马神经元发育的毒性作用

  

  • 收稿日期:2017-09-04 出版日期:2017-10-15 发布日期:2017-10-15

Toxic effect of acrylamide on the development of hippo­campal neurons of weaning rats

Sheng-min Lai, Zi-ting Gu, Meng-meng Zhao, Xi-xia Li, Yu-xin Ma, Li Luo, Jing Liu   

  1. Department of Human Anatomy and Histoembryology, School of Basic Courses, Guangdong Pharmaceutical University, Guangzhou, Guangdong Province, China
  • Received:2017-09-04 Online:2017-10-15 Published:2017-10-15
  • Contact: Jing Liu, Ph.D.,liulq1227@163.com.
  • Supported by:

    This study was supported by the Guangdong Provincial Department of Science and Technology in China, No. 2016A020225007.

摘要:

有关丙烯酰胺的神经毒性在成年动物中已有大量的报道,但关于丙烯酰胺对胚胎期和断乳期动物的神经元发育的研究较少。为验证丙烯酰胺在神经发育期的毒性,作者设计了选择观察神经特异性轴突膜蛋白-生长相关蛋白43和突触前囊泡膜上特异性蛋白-突触素的异常表达,以反映中枢神经系统早期神经元发育的功能紊乱的实验方案。实验将SD母鼠与雄鼠1:1合笼饲养,母鼠从怀孕第6-21天灌胃5,10,20 mg/kg丙烯酰胺染毒,通过进行步态评分来分析母鼠毒性反应情况,每组各产下8只幼鼠,在出生21 d断乳期取材,予尼氏染色观察海马组织学变化,利用免疫组织化学观察包括树突和轴突在内的神经突的状态,再用免疫印迹分析检测神经特异性轴突膜蛋白生长相关蛋白43和突触前囊泡膜上的特异性蛋白突触素表达水平。结果发现母鼠步态Gait评分显著升高,表明丙烯酰胺可造成母鼠运动功能障碍,其产下出生21 d断乳期幼鼠海马神经元数量、生长相关蛋白43和突触素的免疫反应及蛋白的表达均随丙烯酰胺剂量的增加而减少。证明丙烯酰胺对断乳期幼鼠海马神经元的生长与发育均有呈剂量依赖性的毒性作用。

orcid:0000-0002-2416-1162(Jing Liu)

关键词: 神经再生, 丙烯酰胺, 海马, 神经元, 发育毒性, 生长相关蛋白43 , 突触素, 断乳期大鼠, 齿状回, 神经再生, 蛋白, 发育神经生物学

Abstract:

Although numerous studies have examined the neurotoxicity of acrylamide in adult animals, the effects on neuronal development in the embryonic and lactational periods are largely unknown. Thus, we examined the toxicity of acrylamide on neuronal development in the hippocampus of fetal rats during pregnancy. Sprague-Dawley rats were mated with male rats at a 1:1 ratio. Rats were administered 0, 5, 10 or 20 mg/kg acrylamide intragastrically from embryonic days 6–21. The gait scores were examined in pregnant rats in each group to analyze maternal toxicity. Eight weaning rats from each group were also euthanized on postnatal day 21 for follow-up studies. Nissl staining was used to observe histological change in the hippocampus. Immunohistochemistry was conducted to observe the condition of neurites, including dendrites and axons. Western blot assay was used to measure the expression levels of the specific nerve axon membrane protein, growth associated protein 43, and the presynaptic vesicle membrane specific protein, synaptophysin. The gait scores of gravid rats significantly increased, suggesting that acrylamide induced maternal motor dysfunction. The number of neurons, as well as expression of growth associated protein 43 and synaptophysin, was reduced with increasing acrylamide dose in postnatal day 21 weaning rats. These data suggest that acrylamide exerts dose-dependent toxic effects on the growth and development of hippocampal neurons of weaning rats.

Key words: nerve regeneration, acrylamide, hippocampus, neurons, developmental toxicity, growth associated protein 43, synaptophysin, weaning rats, dentate gyrus, protein, developmental neurobiology, neural regeneration