European Journal of Pediatrics

, 167:1169

Repetitive and profound insulin-induced hypoglycemia results in brain damage in newborn rats: an approach to establish an animal model of brain injury induced by neonatal hypoglycemia

  • Dong Zhou
  • Jing Qian
  • Chun-Xi Liu
  • Hong Chang
  • Ruo-Peng Sun
Original Paper

DOI: 10.1007/s00431-007-0653-2

Cite this article as:
Zhou, D., Qian, J., Liu, C. et al. Eur J Pediatr (2008) 167: 1169. doi:10.1007/s00431-007-0653-2


The human neonate is at a higher risk for hypoglycemia-induced neuronal injury than other pediatric and adult patients. Repetitive and profound neonatal hypoglycemia can result in severe neurologic sequelae, of which the mechanisms was not elucidated by hitherto. Moreover, no reliable animal model of brain injury induced by neonatal hypoglycemia is available in order to carry out more research. Therefore, we tried to induce neonatal hypoglycemia in newborn rats by fasting and insulin injection, and then examined the neuronal degeneration after repetitive hypoglycemic insults by Fluoro-Jade B (FJB) staining. Experimental animals were randomly divided into four groups: insulin-treated rats with short hypoglycemia, insulin-treated rats with prolonged hypoglycemia, fasted rats, and control rats. Insulin injection and fasting both could induce consistent hypoglycemia in newborn rats. But from FJB staining results, only in insulin-treated rats with prolonged hypoglycemia could extensive neurodegeneration be detected. We can conclude that FJB staining is a useful method of marking neuronal degeneration in neonatal rats following hypoglycemic brain damage. Repetitive and profound neonatal hypoglycemia can result in extensive neurodegeneration, and it seems that neurons of the cortex, dentate gyrus of the hippocampus, the thalamus, and the hypothalamus are more vulnerable to hypoglycemic insult in newborn rats. Repetitive and profound insulin-induced hypoglycemia in newborn rats can establish a reliable animal model of brain injury resulting from neonatal hypoglycemia.


NeonateHypoglycemiaFluoro-Jade B stainingNeuron



CA1 region of the hippocampus


CA3 region of the hippocampus


Control rats


Dentate gyrus of the hippocampus


Fasted rats


Fluoro-Jade B


Hypodermic injection


Insulin-treated rats with prolonged hypoglycemia


Insulin-treated rats with short hypoglycemia


Postnatal day


Phosphate-buffered saline

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Dong Zhou
    • 1
  • Jing Qian
    • 1
  • Chun-Xi Liu
    • 2
  • Hong Chang
    • 3
  • Ruo-Peng Sun
    • 1
  1. 1.Department of PediatricsQilu Hospital of Shandong UniversityShandongChina
  2. 2.The Key Laboratory of Cardiovascular Remodeling and Function ResearchChinese Ministry of Education and Chinese Ministry of HealthShandongChina
  3. 3.Department of PediatricsMedicine School Hospital of Qingdao UniversityShandongChina