Inhibition of neuronal nitric oxide synthase-mediated activation of poly(ADP-ribose) polymerase in traumatic brain injury: neuroprotection by 3-aminobenzamide

Neuroscience. 2003;121(4):983-90. doi: 10.1016/s0306-4522(03)00482-2.


Focal traumatic injury to the cerebral cortex is associated with early activation of the neuronal isoform of nitric oxide synthase (nNOS), where high concentrations of nitric oxide-derived free radicals elicit extensive DNA damage. Subsequent activation of the nuclear repair enzyme poly(ADP-ribose) polymerase (PARP) causes a severe energy deficit leading to the ultimate demise of affected neurons. Little is known about the temporal relationship of nNOS and PARP activation and the neuroprotective efficacy of their selective blockade in traumatic brain injury. To determine the relationship of nNOS and PARP activation, brain injury was induced by cryogenic lesion to the somatosensory cortex applying a pre-cooled cylinder after trephination for 6 s to the intact dura mater. Pre-treatment with 3-bromo-7-nitroindazole (BrNI; 25 mg/kg, i.p.), and pre- or combined pre- and post-treatment with 3-aminobenzamide (AB; 10 mg/kg (i.c.v.) or 10 mg/kg/h (i.p.)) were used to inhibit nNOS and PARP, respectively. Cold lesion-induced changes in the somatosensory cortex and neuroprotection by BrNI and AB were determined using immunocytochemistry and immunodot-blot for detection of poly(ADP-ribose; PAR), the end-product of PARP activation, and the triphenyltetrazolium-chloride assay to assess lesion volume. PAR immunoreactivity reached its peak 30 min post-lesion and was followed by gradual reduction of PAR immunolabeling. BrNI pre-treatment significantly decreased the lesion-induced PAR concentration in damaged cerebral cortex. Pre-treatment by i.c.v. infusion of AB markedly diminished cortical PAR immunoreactivity and significantly reduced the lesion volume 24 h post-injury. In contrast, i.p. AB treatment remained largely ineffective. In conclusion, our data indicate early activation of PARP after cold lesion that is, at least in part, related to nNOS induction and supports the relevance of nNOS and/or PARP inhibition to therapeutic approaches of traumatic brain injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzamides / pharmacology*
  • Brain Injuries / drug therapy*
  • Brain Injuries / enzymology
  • Brain Injuries / physiopathology
  • Cerebral Infarction / drug therapy
  • Cerebral Infarction / enzymology
  • Cerebral Infarction / prevention & control
  • Cold Temperature / adverse effects
  • Down-Regulation / drug effects
  • Down-Regulation / physiology
  • Drug Interactions / physiology
  • Free Radicals / metabolism
  • Indazoles / pharmacology
  • Male
  • Nerve Degeneration / drug therapy*
  • Nerve Degeneration / enzymology
  • Nerve Degeneration / physiopathology
  • Neuroprotective Agents / pharmacology*
  • Nitric Oxide / metabolism
  • Nitric Oxide Synthase / antagonists & inhibitors*
  • Nitric Oxide Synthase / metabolism
  • Nitric Oxide Synthase Type I
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology
  • Poly(ADP-ribose) Polymerases / metabolism*
  • Rats
  • Rats, Wistar


  • 3-bromo-7-nitroindazole
  • Benzamides
  • Free Radicals
  • Indazoles
  • Neuroprotective Agents
  • Nitric Oxide
  • 3-aminobenzamide
  • Nitric Oxide Synthase
  • Nitric Oxide Synthase Type I
  • Nos1 protein, rat
  • Poly(ADP-ribose) Polymerases