Yamaguchi Y, Okabe K, Liang J, Ohshiro H, Ishihara K, Uchino S, Zhang J L, Hidaka H, Yamada S, Ogawa M
Department of Surgery II, Kumamoto University Medical School, Kumamoto, Japan.
J Surg Res. 2000 Jul;92(1):96-102. doi: 10.1006/jsre.2000.5884.
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.
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.
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.
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.
凝血蛋白酶可能影响白细胞效应功能。缺血/再灌注后凝血级联反应的激活会刺激活化巨噬细胞产生细胞因子。细胞因子诱导的中性粒细胞趋化因子（CINC）在肝缺血/再灌注损伤的病理生理学中可能也很重要。我们研究了选择性因子Xa抑制剂DX-9065a对大鼠肝脏缺血/再灌注后CINC表达的影响。
通过阻断门静脉30分钟诱导大鼠肝脏缺血。在血管夹闭前5分钟静脉注射DX-9065a（9毫克/千克）。采用酶联免疫吸附测定法测量血清CINC浓度。通过Northern印迹分析确定肝脏中CINC mRNA的水平。我们还检测了腹膜巨噬细胞对α-凝血酶或因子Xa的体外CINC产生情况。
血清CINC浓度在再灌注后6小时升高并达到峰值。然而，用DX-9065a预处理动物后，再灌注后CINC的升高明显较小。用DX-9065a预处理还显著降低了缺血/再灌注后肝脏中CINC mRNA的水平。α-凝血酶以及因子Xa均可增强腹膜巨噬细胞的体外CINC产生。
凝血酶和因子Xa刺激巨噬细胞产生CINC。因子Xa的选择性抑制剂DX-9065a可减轻大鼠肝脏缺血/再灌注损伤后中性粒细胞趋化因子的产生。
