Thrombin and factor Xa enhance neutrophil chemoattractant production after ischemia/reperfusion in the rat liver.
BACKGROUND
Clotting proteases may affect leukocyte effector function. Activation of the coagulation cascade after ischemia/reperfusion stimulates cytokine production by activated macrophages. Cytokine-induced neutrophil chemoattractant (CINC) may also be important in the pathophysiology of liver ischemia/reperfusion injury. We investigated the effects of a selective factor Xa inhibitor, DX-9065a, on CINC expression after ischemia/reperfusion in the rat liver.


METHODS
Liver ischemia was induced in rats by occluding the portal vein for 30 min. DX-9065a (9 mg/kg) was injected intravenously 5 min before vascular clamping. Serum CINC concentrations were measured by enzyme-linked immunosorbent assay. Levels of CINC mRNA in the liver were determined by Northern blot analysis. We also examined in vitro CINC production by peritoneal macrophages in response to alpha-thrombin or factor Xa.


RESULTS
Serum CINC concentrations increased and peaked 6 h after reperfusion. However, pretreatment of animals with DX-9065a resulted in significantly smaller increases in CINC after reperfusion. Pretreatment with DX-9065a also significantly reduced CINC mRNA levels in the liver after ischemia/reperfusion. In vitro CINC production by peritoneal macrophages was enhanced by alpha-thrombin, as well as factor Xa.


CONCLUSIONS
Thrombin and factor Xa stimulate CINC production by macrophages. A selective inhibitor of factor Xa, DX-9065a, attenuates neutrophil chemoattractant production after ischemia/reperfusion injury of the rat liver.
Yamaguchi
Y
Y
Department of Surgery II, Kumamoto University Medical School, Kumamoto, Japan.
Okabe
K
K
Liang
J
J
Ohshiro
H
H
Ishihara
K
K
Uchino
S
S
Zhang
J L
JL
Hidaka
H
H
Yamada
S
S
Ogawa
M
M
D000818:Animals; D000925:Anticoagulants / Q000494:pharmacology; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000097:blood / Q000235:genetics*; D015951:Factor Xa / Q000378:metabolism; D065427:Factor Xa Inhibitors; D015870:Gene Expression / Q000502:physiology; D006133:Growth Substances / Q000097:blood / Q000235:genetics*; D006490:Hemostatics / Q000494:pharmacology*; D036341:Intercellular Signaling Peptides and Proteins; D007700:Kinetics; D007728:Kupffer Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000098:blood supply / Q000166:cytology / Q000378:metabolism*; D017737:Macrophages, Peritoneal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D008297:Male; D009281:Naphthalenes / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D011422:Propionates / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology*
C086910:(2S)-2-(4-(((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy)phenyl)-3-(7-amidino-2-naphtyl)propanoic acid; D000925:Anticoagulants; D019743:Chemokines, CXC; D002630:Chemotactic Factors; D065427:Factor Xa Inhibitors; D006133:Growth Substances; D006490:Hemostatics; D036341:Intercellular Signaling Peptides and Proteins; D009281:Naphthalenes; D011422:Propionates; D012333:RNA, Messenger; D013917:Thrombin; D015951:Factor Xa