Furuno T, Mitsuyama T, Hidaka K, Tanaka T, Hara N
Research Institute for Diseases of the Chest, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
Int Arch Allergy Immunol. 1997 Mar;112(3):262-9. doi: 10.1159/000237464.
Neutrophils are thought to play a key role in tissue injury. We investigated the role of human neutrophil-derived elastase in the induction of injury to human pulmonary artery endothelial cells. Incubation of endothelial cells with neutrophils increased the release of lactate dehydrogenase activity, thrombomodulin, and preloaded fura-2 from endothelial cells, indicating that neutrophils induce endothelial cell injury. Attachment alone of neutrophils to endothelial cells appeared to induce activation because elastase release and N-formyl-mentionyl-leucyl-phenylalanine (fMLP)-induced superoxide (O2) production from neutrophils incubated with endothelial cells were greater than from neutrophils only. When endothelial cell were incubated with neutrophils stimulated by fMLP or phorbol myristate acetate, the amount of elastase in the medium and endothelial cell damage was further enhanced. However, when neutrophils were blocked from direct attachment to endothelial cells using a membrane filter, endothelial cell damage was ameliorated, while exogenous neutrophil elastase and medium containing neutrophil-released elastase did not induce endothelial cell injury. An inhibitor of neutrophil elastase, ONO-5046 Na, as well as erythromycin, which reduces neutrophil-derived elastase, dramatically inhibited neutrophil-induced endothelial cell injury. Superoxide dismutase (SOD) partially inhibited injury. Injury was completely inhibited by treatment with a combination of ONO-5046 Na and SOD. These results suggest that attachment of neutrophils to endothelial cells is important for endothelial cell damage and that neutrophil-derived elastase plays an important role in endothelial cell injury in combination with O2. In addition, ONO-5046 Na and erythromycin may be useful in treating diseases worsened by excessive neutrophil activity.
中性粒细胞被认为在组织损伤中起关键作用。我们研究了人中性粒细胞衍生的弹性蛋白酶在诱导人肺动脉内皮细胞损伤中的作用。内皮细胞与中性粒细胞共孵育增加了乳酸脱氢酶活性、血栓调节蛋白以及内皮细胞中预加载的fura-2的释放，表明中性粒细胞可诱导内皮细胞损伤。中性粒细胞仅与内皮细胞附着似乎就可诱导激活，因为与内皮细胞共孵育的中性粒细胞的弹性蛋白酶释放以及N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（fMLP）诱导的超氧化物（O2）产生量比仅中性粒细胞的情况更多。当内皮细胞与经fMLP或佛波酯刺激的中性粒细胞共孵育时，培养基中的弹性蛋白酶量和内皮细胞损伤进一步加剧。然而，当使用膜滤器阻止中性粒细胞直接附着于内皮细胞时，内皮细胞损伤得到改善，而外源性中性粒细胞弹性蛋白酶和含有中性粒细胞释放的弹性蛋白酶的培养基并未诱导内皮细胞损伤。中性粒细胞弹性蛋白酶抑制剂ONO-5046 Na以及可减少中性粒细胞衍生弹性蛋白酶的红霉素可显著抑制中性粒细胞诱导的内皮细胞损伤。超氧化物歧化酶（SOD）可部分抑制损伤。用ONO-5046 Na和SOD联合处理可完全抑制损伤。这些结果表明，中性粒细胞与内皮细胞的附着对于内皮细胞损伤很重要，并且中性粒细胞衍生的弹性蛋白酶与O2共同在内皮细胞损伤中起重要作用。此外，ONO-5046 Na和红霉素可能有助于治疗因中性粒细胞活性过高而恶化的疾病。
