PMID- 19108900
OWN - NLM
STAT- MEDLINE
DCOM- 20090818
LR  - 20171116
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 129
IP  - 3-4
DP  - 2009 Jun 15
TI  - Endotoxin-induced activation of equine digital vein endothelial cells: role of 
      p38 MAPK.
PG  - 174-80
LID - 10.1016/j.vetimm.2008.11.008 [doi]
AB  - The endothelium plays a major role in the pathogenesis of endotoxemia. Binding of 
      endotoxin (lipopolysaccharide; LPS) to endothelium initiates a range of 
      signalling events, including activation of p38 mitogen activated protein kinases 
      (MAPKs) that are involved in the initiation of inflammatory responses. In the 
      present study we have examined whether clinically relevant concentrations of LPS 
      can activate p38 MAPK in equine endothelial cells and have investigated the role 
      of the kinase in neutrophil adhesion and mediator release. Cultured equine 
      digital vein endothelial cells (EDVEC) were exposed to LPS and phosphorylation of 
      p38 MAPK was assessed by Western blotting using phospho-specific antibodies. 
      Neutrophil adhesion was quantified by assaying myeloperoxidase and the release of 
      prostacyclin (PGI(2)) was measured by radioimmunoassay of its stable breakdown 
      product 6-keto-PGF(1alpha). The effects of the p38 MAPK inhibitors, SB203580 and 
      PD169316, on neutrophil adhesion and 6-keto-PGF(1alpha) release were examined, as 
      was the effect of an anti-E-selectin antibody on neutrophil adhesion to 
      LPS-activated EDVEC. LPS treatment significantly increased p38 MAPK 
      phosphorylation, which was maximal after a 1h LPS incubation using a 
      concentration of 10(-5)g/ml (EC(50) = 2 x 10(-7)g/ml). Neutrophil adhesion to 
      LPS-stimulated endothelial cells (maximal at 10(-6)g/ml; EC(50) = 6 x 
      10(-10)g/ml) was significantly inhibited in the presence of p38 MAPK inhibitors 
      (reduced from a maximum of 33+/-6% to 13+/-4% adhesion at 10(-6)M SB203580 and to 
      21+/-4% adhesion at 10(-6) M PD169316), or an anti-E-selectin antibody (reduced 
      from 17+/-1% adhesion to 6+/-1% adhesion). 6-keto-PGF(1alpha) release was 
      increased in a concentration-dependent manner following LPS exposure (maximal at 
      10(-6)g/ml; EC(50) = 1 x 10(-9)g/ml), and was significantly inhibited by p38 MAPK 
      blockade (reduced from 1.6+/-0.3 x 10(-9)g/ml to 4+/-1 x 10(-10)g/ml and 4+/-2 x 
      10(-10)g/ml with 10(-6) M SB203580 or 10(-6) M PD169316, respectively). This 
      study has demonstrated that clinically relevant concentrations of LPS activate 
      p38 MAPK in equine endothelial cells and that both neutrophil adhesion to 
      LPS-activated EDVEC and PGI(2) release are dependent upon p38 MAPK 
      phosphorylation. These results reveal a key role for p38 MAPK in the response of 
      the endothelium to LPS and suggest that inhibition of this kinase may reduce 
      inflammatory events in the endotoxemic horse.
FAU - Brooks, Andrew C
AU  - Brooks AC
AD  - Department of Veterinary Basic Sciences, The Royal Veterinary College, Hawkshead 
      Lane, North Mymms, Hertfordshire AL9 7TA, UK. abrooks@rvc.ac.uk
FAU - Menzies-Gow, Nicola J
AU  - Menzies-Gow NJ
FAU - Wheeler-Jones, Caroline
AU  - Wheeler-Jones C
FAU - Bailey, Simon R
AU  - Bailey SR
FAU - Cunningham, Fiona M
AU  - Cunningham FM
FAU - Elliott, Jonathan
AU  - Elliott J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20081107
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Imidazoles)
RN  - 0 (Lipopolysaccharides)
RN  - 0 (Pyridines)
RN  - DCR9Z582X0 (Epoprostenol)
RN  - EC 2.7.11.24 (p38 Mitogen-Activated Protein Kinases)
RN  - GX3Y2V80CV (2-(4-nitrophenyl)-4-(4-fluorophenyl)-5-(4-pyridinyl)-1H-imidazole)
RN  - OU13V1EYWQ (SB 203580)
SB  - IM
MH  - Animals
MH  - Cell Adhesion
MH  - Cells, Cultured
MH  - Endothelial Cells/*drug effects/physiology
MH  - Enzyme Inhibitors/pharmacology
MH  - Epoprostenol/metabolism
MH  - Hoof and Claw/*blood supply
MH  - Horses/*physiology
MH  - Imidazoles/pharmacology
MH  - Lipopolysaccharides/*pharmacology
MH  - Neutrophils/physiology
MH  - Phosphorylation
MH  - Pyridines/pharmacology
MH  - Veins/drug effects/physiology
MH  - p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors/*metabolism
EDAT- 2008/12/26 09:00
MHDA- 2009/08/19 09:00
CRDT- 2008/12/26 09:00
PHST- 2008/12/26 09:00 [entrez]
PHST- 2008/12/26 09:00 [pubmed]
PHST- 2009/08/19 09:00 [medline]
AID - S0165-2427(08)00734-4 [pii]
AID - 10.1016/j.vetimm.2008.11.008 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2009 Jun 15;129(3-4):174-80. doi: 
      10.1016/j.vetimm.2008.11.008. Epub 2008 Nov 7.