Nakatani K, Takeshita S, Tsujimoto H, Kawamura Y, Sekine I
Department of Pediatrics, National Defense Medical College, Tokorozawa, Saitama, Japan.
J Leukoc Biol. 2001 Feb;69(2):241-7.
To investigate the inhibitory effect of serine protease inhibitors (SPI) on neutrophil-mediated endothelial cell (EC) injury, we analyzed the in vitro cytotoxicity of radiolabeled human umbilical vein EC (HUVEC) mediated by neutrophils in the presence of SPI. The EC injury was inhibited dose-dependently by urinary trypsin inhibitor (ulinastatin, UTI) and ONO-5046, which have the ability to inactivate neutrophil elastase, but not by gabexate mesilate, nafamostat mesilate, aprotinin, and argatroban, which have no ability to inactivate neutrophil elastase. In addition, when UTI and ONO-5046 were added to the tumor necrosis factor alpha-primed neutrophils alone, they showed a dose-dependent inhibition of the intracellular elastase activity, but the other SPI did not, for either flow cytometry or confocal microscopy. Therefore, UTI and ONO-5046 may protect EC against the neutrophil-mediated injury not only by inactivating the extracellular elastase secreted by neutrophils, but also by acting directly on neutrophils and suppressing the production and secretion of activated elastase from them.
为研究丝氨酸蛋白酶抑制剂（SPI）对中性粒细胞介导的内皮细胞（EC）损伤的抑制作用，我们分析了在SPI存在的情况下，放射性标记的人脐静脉内皮细胞（HUVEC）由中性粒细胞介导的体外细胞毒性。尿胰蛋白酶抑制剂（乌司他丁，UTI）和ONO - 5046对EC损伤具有剂量依赖性抑制作用，它们具有使中性粒细胞弹性蛋白酶失活的能力，而甲磺酸加贝酯、甲磺酸萘莫司他、抑肽酶和阿加曲班则无此作用，它们没有使中性粒细胞弹性蛋白酶失活的能力。此外，当单独将UTI和ONO - 5046添加到肿瘤坏死因子α预处理的中性粒细胞中时，通过流式细胞术或共聚焦显微镜观察，它们均显示出对细胞内弹性蛋白酶活性的剂量依赖性抑制作用，但其他SPI则没有此作用。因此，UTI和ONO - 5046可能不仅通过使中性粒细胞分泌的细胞外弹性蛋白酶失活，而且通过直接作用于中性粒细胞并抑制其活化弹性蛋白酶的产生和分泌来保护EC免受中性粒细胞介导的损伤。
