Multiple selectin blockade with a small molecule inhibitor downregulates liver chemokine expression and neutrophil infiltration after hemorrhagic shock.
BACKGROUND
The purpose of this study was to investigate the regulatory effect of a small molecule selectin inhibitor in the liver by examining the functional, structural, and survival response of animals subjected to hemorrhagic shock and to determine the liver infiltration of neutrophils and the regulation of chemokine expression. Selectins play an important role in the development of the lesions associated with ischemia/reperfusion and hemorrhagic shock. Blocking individually the selectin family of adhesion molecules with monoclonal antibodies has resulted in better organ function and survival. To our knowledge, there are no studies demonstrating the beneficial effect of multiple selectin blockade with a small molecule inhibitor under conditions of hemorrhagic shock.


METHODS
Forty-eight Sprague-Dawley rats were subjected to hemorrhagic shock. Three groups of animals were included (n = 16/group), i.e., the sham, control, and treated groups, which received a small molecule selectin inhibitor (TBC-1269) at 25 mg/kg body weight after the bleeding began. The following parameters were evaluated: fluid requirements during resuscitation, liver injury tests (aspartate aminotransferase, alanine aminotransferase), liver histology and myeloperoxidase, and macrophage inflammatory protein-2 mRNA and cytokine-induced neutrophil chemoattractant mRNA in liver tissue, and animal survival at 3 days. Statistical analysis included Student's t test and analysis of variance when indicated.


RESULTS
Significant improvement in liver function and histology was noted in the treated group. Survival was also improved, although it is not known whether liver failure was the most proximate cause of lethality. Infiltration of neutrophils, measured by tissue myeloperoxidase, was significantly decreased in livers of treated animals. No significant changes were noted in fluid requirements. The small molecule selectin inhibitor group showed a down-regulating effect on liver macrophage inflammatory protein-2 and cytokine-induced neutrophil chemoattractant mRNA expression associated with less accumulation of neutrophils in the liver.


CONCLUSIONS
This study supports the role that selectins play in the pathogenesis of hemorrhagic shock. The mechanism of protection seen after multiple selectin blockade (TBC-1269) centered, in part, around the infiltration of liver neutrophils, probably dependent on the induction of macrophage inflammatory protein-2 and cytokine-induced neutrophil chemoattractant mRNA expression in liver tissue.
Borgess Research Institute, Surgery Research Sciences and Molecular Biology, Kalamazoo, Michigan, USA.
J R
JR
Ramos-Kelly
L H
LH
Toledo-Pereyra
J
J
Jordan
F
F
Rivera-Chavez
T
T
Rohs
M
M
Holevar
R A
RA
Dixon
E
E
Yun
P A
PA
Ward
D000704:Analysis of Variance; D000818:Animals; D001713:Biphenyl Compounds / Q000494:pharmacology*; D015152:Blotting, Northern; D018925:Chemokines / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D015536:Down-Regulation; D008099:Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism* / Q000473:pathology; D008297:Male; D008358:Mannose / Q000031:analogs & derivatives; D008362:Mannosides / Q000494:pharmacology*; D009504:Neutrophils / Q000378:metabolism; D009195:Peroxidase / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D019087:Selectins / Q000502:physiology*; D012771:Shock, Hemorrhagic / Q000378:metabolism* / Q000473:pathology
D001713:Biphenyl Compounds; D018925:Chemokines; D008362:Mannosides; D012333:RNA, Messenger; D019087:Selectins; C108700:bimosiamose disodium; D009195:Peroxidase; D008358:Mannose