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Inflammation Research

, Volume 56, Issue 4, pp 154–161 | Cite as

Endotoxin-induced activation of equine platelets: evidence for direct activation of p38 MAPK pathways and vasoactive mediator production

  • A. C. BrooksEmail author
  • N. J. Menzies-Gow
  • C. Wheeler-Jones
  • S. R. Bailey
  • F. M. Cunningham
  • J. Elliott
Original Research Paper

Abstract.

Objective and design:

The aim of this study was to determine the effects of endotoxin on p38 MAPK activation in equine platelets and leukocytes in vivo and in vitro and its role in thromboxane (Tx) production with reference to equine endotoxaemia.

Methods:

Six adult Thoroughbred horses were used for in vivo infusion studies and separate in vitro studies. For in vivo studies, following collection of a pre-infusion sample, horses were infused with E. Coli O55:B5 LPS (30 ng/kg; 30 min) during and after which platelets were harvested. For in vitro studies isolated platelets and leukocytes were exposed to LPS (10 pg/ml–1 μg/ml). p38 MAPK activity was assessed by SDS-PAGE followed by immunoblotting. TxA2 release was measured by radioimmunoassay.

Results:

LPS infusion caused increased phospho-p38 MAPK in equine platelets and leukocytes (1492 ± 486 % and 83 ± 45 above basal, respectively) from 10 min after the start of the infusion, which returned to basal by 60 min. In vitro, platelets were 1,000 times more sensitive to LPS than leukocytes in terms of both TxA2 production (EC50 66 pg/ml versus 110 ng/ml, respectively) and p38 MAPK phosphorylation (EC50 11.1 ± 2 pg/ml versus 14.8 ± 4 ng/ml, respectively). p38 MAPK inhibitors SB203580 and PD169316 attenuated LPS-induced TxA2 release in platelets, but not leukocytes.

Conclusions:

In vivo, LPS stimulates TxA2 production and p38 MAPK phosphorylation in equine platelets and leukocytes at a concentration within a similar range to those reported in clinical endotoxaemia. These data suggest that LPS-induced eicosanoid production in the early phase of clinical endotoxaemia may involve direct effects of LPS upon platelets, mediated via activation of p38 MAPK.

Keywords

Cell signaling Endotoxin Platelets 

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Copyright information

© Birkhäuser Verlag, Basel 2007

Authors and Affiliations

  • A. C. Brooks
    • 1
    Email author
  • N. J. Menzies-Gow
    • 2
  • C. Wheeler-Jones
    • 1
  • S. R. Bailey
    • 1
  • F. M. Cunningham
    • 1
  • J. Elliott
    • 1
  1. 1.Departments of Veterinary Basic SciencesThe Royal Veterinary CollegeHertfordshireUK
  2. 2.Veterinary Clinical SciencesRoyal Veterinary CollegeHertfordshireUK

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