Early nuclear factor-kB activation and inducible nitric oxide synthase expression in injured spinal cord neurons correlating with a diffuse reduction of constitutive nitric oxide synthase activity

Sabrina Ceccariglia, Liverana Lauretti, Aurora Del Fa' Gangitano, Carlo Gangitano, Massimo Miscusi, Florian Ebner, Marta Menegazzi, Sofia Mariotto, Luigi Berra, Paolo Missori, Roberto Delfini, Hisamori Suzuki

Risultato della ricerca: Contributo in rivistaArticolo in rivista

Abstract

OBJECT: Because of toxicity at high concentrations, nitric oxide (NO) contributes to spinal cord injury (SCI) secondary lesions. At low concentrations NO modulates nuclear factor-kappaB (NF-kappaB) activation. The authors investigated the activity of neuronal and endothelial NO synthase (nNOS and eNOS) to determine correlations with NF-kappaB activation and inducible NOS (iNOS) expression soon after SCI. METHODS: In 48 adult male Wistar rats clip-based (50 g/mm2/10 seconds) SCI was induced, and spinal cords were removed at different intervals for the following evaluations: 1) assaying specific activity of nNOS and eNOS; 2) electrophoresis mobility shift assay for activated NF-kappaB; 3) Northern blotting for iNOS; 4) immunohistochemistry for iNOS and NF-kappaB; and 5) immunofluorescence for iNOS and NF-kappaB. At 15 minutes postinjury, eNOS activity decreased significantly (p < 0.001), as did nNOS activity at 1 hour compared with these levels in control animals and rats killed at 15 and 30 minutes after SCI (p < 0.001). Basal NF-kappaB levels were variable in controls and at 15 and 30 minutes after injury. One hour postinjury, NF-kappaB activation was diffuse. Inducible NOS messenger RNA localized diffusely, peaking 6 hours after injury and remaining stable until 24 hours postinjury. Immunohistochemical analysis showed diffuse iNOS and NF-kappaB staining, especially in neurons inside and around the lesion. Immunofluorescence demonstrated that injured neurons were a source of NF-kappaB and iNOS soon after injury. CONCLUSIONS: Both nNOS and eNOS exhibited different regulation and roles soon after injury: nNOS correlated with NF-kappaB activation, whereas eNOS may have participated in vascular changes of the injured spinal cord. Neurons seemed to play a pivotal role in modulating and amplifying the inflammatory response in the injured spinal cord.
Lingua originaleEnglish
pagine (da-a)485-493
Numero di pagine9
RivistaJOURNAL OF NEUROSURGERY. SPINE
Stato di pubblicazionePubblicato - 2006

Keywords

  • constitutive nitric oxide synthase
  • inducible nitric oxide synthase
  • nitric oxid
  • nuclear factor-kB
  • spinal cord iniury

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