Allopregnanolone attenuates N-methyl-D-aspartate-induced excitotoxicity and apoptosis in the human NT2 cell line in culture

Neurosci Lett. 2002 Aug 2;328(1):33-6. doi: 10.1016/s0304-3940(02)00448-2.

Abstract

Progesterone modulates gamma-aminobutyric acid and excitatory amino acid neurotransmitter systems and has neuroprotective properties in models of hypoxia-ischemia. This study examined the in vitro effects of allopregnanolone, the active progesterone metabolite, in models of N-methyl-D-aspartate (NMDA)-induced necrosis and apoptosis. Cultured NT2 neurons were exposed to 1 mM NMDA. Lactate dehydrogenase (LDH) release was measured 24 h later. NMDA at a concentration of 1 mM produced a 39 +/- 19% release of total LDH. Exposure to 10 microM allopregnanolone prior to NMDA exposure reduced LDH release by 51% (P = 0.0028). NMDA stimulated apoptotic cell changes defined by terminal dUTP nick-end labeling (TUNEL) and 5,5', 6,6'-tetrachloro-1,1,3,3'-tetra ethlybenzimidazolycarbocyanide iodide staining were reduced to baseline values by both 10 microM allopregnanolone and 100 microM MK-801. Pretreatment with allopregnanolone (0-10 microM) reduced the percentage of TUNEL-positive cells in a dose-dependent manner (EC(50) = 2.7 +/- 0.1 nM). Physiologic concentrations of allopregnanolone provided protection against both necrotic and apoptotic injury induced by NMDA excitotoxicity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoptosis / drug effects
  • Apoptosis / physiology*
  • Asphyxia Neonatorum / metabolism*
  • Asphyxia Neonatorum / physiopathology
  • Benzimidazoles
  • Carbocyanines
  • Cell Count
  • Cell Survival / drug effects
  • Cell Survival / physiology*
  • Dizocilpine Maleate / pharmacology
  • Dose-Response Relationship, Drug
  • Excitatory Amino Acid Agonists / pharmacology
  • Excitatory Amino Acid Antagonists / pharmacology
  • Female
  • Fluorescent Dyes
  • Humans
  • Hypoxia-Ischemia, Brain / drug therapy
  • Hypoxia-Ischemia, Brain / metabolism*
  • Hypoxia-Ischemia, Brain / physiopathology
  • In Situ Nick-End Labeling
  • Infant, Newborn
  • L-Lactate Dehydrogenase / drug effects
  • L-Lactate Dehydrogenase / metabolism
  • Membrane Potentials / drug effects
  • Mitochondria / drug effects
  • Mitochondria / metabolism
  • N-Methylaspartate / pharmacology
  • Neurons / drug effects
  • Neurons / metabolism*
  • Neuroprotective Agents / metabolism
  • Neuroprotective Agents / therapeutic use*
  • Neurotoxins / pharmacology
  • Pregnancy
  • Pregnanolone / pharmacology
  • Pregnanolone / therapeutic use
  • Progesterone / metabolism
  • Progesterone / therapeutic use*
  • Tumor Cells, Cultured

Substances

  • Benzimidazoles
  • Carbocyanines
  • Excitatory Amino Acid Agonists
  • Excitatory Amino Acid Antagonists
  • Fluorescent Dyes
  • Neuroprotective Agents
  • Neurotoxins
  • 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine
  • Progesterone
  • N-Methylaspartate
  • Dizocilpine Maleate
  • Pregnanolone
  • L-Lactate Dehydrogenase