Highlights
- •Cilostazol increased p-AMPK- and p-p38 that followed by HO-1 induction.
- •Cilostazol reduced HMGB1 and PAI-1 in septic mice in a ZnPPIX-sensitive manner.
- •Cilostazol increased the survival of endotoxemic mice.
Abstract
Introduction
Inflammation and coagulation play important roles in the pathogenesis of sepsis. Anticoagulants
with anti-inflammatory action draw attention as therapeutic agent in sepsis.
Objective
Whether cilostazol (6-[4-(1-cyclohexyl-1H-tetrazol-5-yl) butoxy]-3,4-dihydro-2-(1H)-quinolinone),
anticoagulant, protects mice against sepsis and underlying mechanism(s) were investigated.
Methods
Induction of heme oxygenase (HO)-1 protein, phosphorylation of 5' adenosine monophosphate-activated
protein kinase (AMPK), nuclear factor kappa-light-chain-enhancer of activated B cells
(NF-κB) luciferase activity, and release of high mobility group box 1 (HMGB1) were
analyzed using signal inhibitors and transfection techniques. Survival and organ damage
were compared in septic mice with and without cilostazol.
Results
In RAW264.7 cells, cilostazol increased phosphorylation of AMPK which was followed
by HO-1 induction. Lipopolysaccharide (LPS)-activated HMGB1 release was reduced by
cilostazol which was reversed by both SB203580 and silencing of HO-1 or AMPK RNA.
Interestingly, silencing AMPK reduced HO-1 expression, whereas silencing HO-1 did
not affect p-AMPK by cilostazol. Both compound C and zinc protoporphyrin IX (ZnPPIX)
antagonized inhibitory effect of HMGB1 by cilostazol. Cilostazol inhibited NF-κB luciferase
activity which was antagonized by SB203580. Finally, the administration of cilostazol
increased the survival of endotoxemic mice but failed to do so when co-treated with
rHMGB1. Cilostazol reduced circulating HMGB1, plasminogen activator inhibitor-1 (PAI-1)
levels, organ damages and protein expression of PAI-1 in lung tissues of CLP-septic
mice, which were antagonized by ZnPPIX.
Conclusion
These findings suggest that HMGB1 can be a target molecule of cilostazol by 1) AMPK
activation, and 2) induction of HO-1 by p38 MAPK and AMPK. Therefore, cilostazol may
be useful for treatment of sepsis.
Graphical Abstract
Graphical Abstract
Abbreviations:
AMPK (5' adenosine monophosphate-activated protein kinase), ATCC (American Type Culture Collection), CO (Carbon monoxide), CLP (Cecal ligation and puncture), Cilostazol ((6-[4-(1-cyclohexyl-1H-tetrazol-5-yl) butoxy]-3,4-dihydro-2-(1H)-quinolinone)), DMEM (Dulbecco’s modified Eagle’s medium), ECs (Endothelial cells), ECL (Enhanced chemiluminescence), FBS (Fetal bovine serum), HO-1 (Heme oxygenase-1), HMGB1 (High-mobility group box 1), IFN-γ (Interferon-gamma), IL-1 (Interleukin-1), LPS (Lipopolysaccharide), NF-κB (Nuclear factor kappa-light-chain-enhancer of activated B cells), rHMGB1 (Recombinant human HMGB1), ZnPPIX (Zinc protoporphyrin IX)Keywords
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Article info
Publication history
Published online: June 15, 2015
Accepted:
June 14,
2015
Received in revised form:
May 11,
2015
Received:
February 6,
2015
Identification
Copyright
© 2015 Elsevier Ltd. Published by Elsevier Inc. All rights reserved.
