PMID- 11474119
OWN - NLM
STAT- MEDLINE
DCOM- 20010816
LR  - 20180821
IS  - 1076-1551 (Print)
IS  - 1528-3658 (Electronic)
IS  - 1076-1551 (Linking)
VI  - 6
IP  - 12
DP  - 2000 Dec
TI  - Tumor necrosis factor and reactive oxygen species cooperative cytotoxicity is 
      mediated via inhibition of NF-kappaB.
PG  - 1028-41
AB  - BACKGROUND: Tumor necrosis factor alpha (TNFalpha) plays a key role in 
      pathogenesis of brain injury. However, TNFalpha exhibits no cytotoxicity in 
      primary cultures of brain cells. This discrepancy suggests that other pathogenic 
      stimuli that exist in the setting of brain injury precipitate TNFalpha 
      cytotoxicity. The hypothesis was tested that reactive oxygen species (ROS), that 
      are released early after brain injury, act synergistically with TNFalpha in 
      causing cell death. MATERIALS AND METHODS: Cultured human and rat brain capillary 
      endothelial cells (RBEC), and cortical astrocytes were treated with TNFalpha 
      alone or together with different doses of H2O2, and apoptotic cell death and DNA 
      fragmentation were measured by means of 3'-OH-terminal deoxynucleotidyl 
      transferase-mediated dUTP nick end labeling (TUNEL) and Hoechst fluorescence 
      assay, respectively. The effect of H2O2 on TNFalpha-induced activation of nuclear 
      factor kappa B (NF-kappaB) was measured by Western blots of cytoplasmic and 
      nuclear extracts of RBEC using anti-inhibitor of NF-kappaB (IkappaB) and anti-p65 
      subunit of NF-kappaB antibodies. Nuclear translocation of NF-kappaB was 
      investigated by immunofluorescent staining of RBEC with anti-p65 antibodies. 
      RESULTS: TNFalpha alone had no cytotoxic effect in brain endothelial cells and 
      astrocytes at concentrations up to 100 ng/ml. Co-treatment with 5-10 microM of 
      H2O2 caused a two-fold increase in the number of apoptotic cells 24 hr later. 
      Similar doses (1-3 microM) of H2O2 initiated early DNA fragmentation. H2O2 
      inhibited TNFalpha-induced accumulation of p65 in the nucleus, although it had no 
      effect on degradation of the IkappaB in cytoplasm. Immunostaining confirmed that 
      H2O2 inhibited p65 transport to the nucleus. CONCLUSIONS: Reactive oxygen species 
      could act synergistically with TNFalpha in causing cytotoxicity via inhibition of 
      a cytoprotective branch of TNFalpha signaling pathways, which starts with 
      NF-kappaB activation.
FAU - Ginis, I
AU  - Ginis I
AD  - Stroke Branch, National Institute of Neurological Disorders and Stroke, NIH, 
      Bethesda, MD 20892-4128, USA. ginisi@ninds.nih.gov
FAU - Hallenbeck, J M
AU  - Hallenbeck JM
FAU - Liu, J
AU  - Liu J
FAU - Spatz, M
AU  - Spatz M
FAU - Jaiswal, R
AU  - Jaiswal R
FAU - Shohami, E
AU  - Shohami E
LA  - eng
PT  - Journal Article
PL  - England
TA  - Mol Med
JT  - Molecular medicine (Cambridge, Mass.)
JID - 9501023
RN  - 0 (NF-kappa B)
RN  - 0 (Reactive Oxygen Species)
RN  - 0 (Transcription Factor RelA)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - BBX060AN9V (Hydrogen Peroxide)
SB  - IM
MH  - Animals
MH  - Apoptosis
MH  - Astrocytes/cytology/metabolism
MH  - Blotting, Western
MH  - Brain/*metabolism
MH  - Capillaries/metabolism
MH  - Cell Nucleus/metabolism
MH  - Cells, Cultured
MH  - Cytosol/metabolism
MH  - DNA Fragmentation
MH  - Dose-Response Relationship, Drug
MH  - Endothelium, Vascular/cytology/metabolism
MH  - Enzyme Activation
MH  - Humans
MH  - Hydrogen Peroxide/pharmacology
MH  - In Situ Nick-End Labeling
MH  - Microscopy, Fluorescence
MH  - NF-kappa B/*metabolism/*physiology
MH  - Rats
MH  - Reactive Oxygen Species/*metabolism
MH  - Time Factors
MH  - Transcription Factor RelA
MH  - Tumor Necrosis Factor-alpha/*metabolism
PMC - PMC1949928
EDAT- 2001/07/28 10:00
MHDA- 2001/08/17 10:01
PMCR- 2000/12/01
CRDT- 2001/07/28 10:00
PHST- 2001/07/28 10:00 [pubmed]
PHST- 2001/08/17 10:01 [medline]
PHST- 2001/07/28 10:00 [entrez]
PHST- 2000/12/01 00:00 [pmc-release]
AID - S1528365800210288 [pii]
PST - ppublish
SO  - Mol Med. 2000 Dec;6(12):1028-41.