TPEN Attenuates Neural Autophagy Induced by Synaptically-released Zinc Translocation and Improves Histological Outcomes after Traumatic Brain Injury in Rats

Ann Clin Lab Sci. 2018 Jul;48(4):446-452.

Abstract

Objectives: Previous studies have demonstrated that the inhibition of autophagy could reduce traumatic brain injury (TBI)-induced cell injury and attenuate behavioural outcomes. Meanwhile, synaptically released zinc translocation is found in the hippocampus of rats, and excessive release of chelatable zinc from excitatory synaptic vesicles is involved in the pathophysiological processes of TBI. We speculated that the release of zinc is closely connected with autophagy and that treatment with the zinc chelator N,N,N',N'-tetrakis-(2-pyridylmethyl) ethylenediamine (TPEN) could attenuate autophagy and thus improve histologic outcomes after TBI in rats.

Methods: Rats were randomly divided into three groups: sham group, CCI+vehicle group and CCI+TPEN group. TPEN (20 mg/kg/12 h, i.p) or vehicle (DMSO) was injected into the rats immediately after TBI, then given twice a day until the animals were sacrificed. Morphological changes associated with autophagy were tested by TEM. Zinc autometallography staining was used to evaluate chelatable zinc accumulation. Western blot analysis was used to detect protein expression.

Results: In our study, we found that treatment with TPEN reduced zinc translocation, reduced excitotoxicity and also partially reduced levels of LC3-II, Beclin1 and the ratio of Beclin-1/Bcl-2 in injured hippocampal neurons of rats, partially improving histologic outcomes after TBI.

Conclusion: These data show that the zinc chelator TPEN plays a protective role in TBI, suggesting the possible involvement of autophagy.

Keywords: TPEN; autophagy; traumatic brain injury; zinc translocation.

MeSH terms

  • Animals
  • Autophagy / drug effects*
  • Beclin-1 / metabolism
  • Brain Injuries, Traumatic / drug therapy*
  • Brain Injuries, Traumatic / metabolism
  • Brain Injuries, Traumatic / pathology*
  • Chelating Agents / pharmacology
  • Chelating Agents / therapeutic use
  • Chronic Disease
  • Constriction, Pathologic
  • Ethylenediamines / pharmacology
  • Ethylenediamines / therapeutic use*
  • Hippocampus / pathology
  • Male
  • Microtubule-Associated Proteins / metabolism
  • Neurons / drug effects
  • Neurons / metabolism
  • Neurons / pathology*
  • Neurons / ultrastructure
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Rats, Sprague-Dawley
  • Synapses / drug effects
  • Synapses / metabolism
  • Synapses / pathology*
  • Synapses / ultrastructure
  • Treatment Outcome
  • Zinc / therapeutic use*

Substances

  • Beclin-1
  • Becn1 protein, rat
  • Chelating Agents
  • Ethylenediamines
  • LC3 protein, rat
  • Microtubule-Associated Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Zinc
  • N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine