NEUTROPHIL-MEDIATED INJURY TO ENDOTHELIAL-CELLS - ENHANCEMENT BY ENDOTOXIN AND ESSENTIAL ROLE OF NEUTROPHIL ELASTASE
NEUTROPHIL-MEDIATED INJURY TO ENDOTHELIAL-CELLS - ENHANCEMENT BY ENDOTOXIN AND ESSENTIAL ROLE OF NEUTROPHIL ELASTASE
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DOI：
10.1172/jci112426
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发表时间：
1986-04-01
影响因子：
15.9
通讯作者：
WORTHEN, GS
中科院分区：
文献类型：
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作者：
SMEDLY, LA;TONNESEN, MG;WORTHEN, GS
中性粒细胞被认为是血管损伤的重要介质，尤其是内毒素血症后。本研究探讨了中性粒细胞介导的体外人微血管内皮细胞损伤。我们发现，在4小时的实验中，由甲酰基-蛋氨酸-乙酰基-苯丙氨酸（FMLP）、补体片段C5a或脂多糖（LPS） （1-1,000 ng/ml）单独刺激的中性粒细胞产生最小的内皮损伤。相比之下，中性粒细胞与内皮细胞在低浓度LPS （1-10 ng/ml）下孵育，FMLP或C5a会刺激中性粒细胞产生明显的内皮损伤。LPS浓度为100 ng/ml， FMLP浓度为10-7 M时损伤最大。用LPS预处理中性粒细胞导致类似程度的损伤，表明LPS的作用主要是对中性粒细胞。lps暴露、fmlp刺激的中性粒细胞对内皮细胞损伤的时间过程与添加纯化的人中性粒细胞弹性酶诱导的时间过程相似，而与过氧化氢（H2O2）诱导的时间过程不同。此外，中性粒细胞介导的损伤不受多种氧自由基清除剂的抑制，并且发生在慢性肉芽肿病患者的中性粒细胞中，其不产生H2O2。相比之下，特异性丝氨酸弹性酶抑制剂甲氧基-琥珀酰-丙酰-丙酰-戊酰-氯甲基酮对中性粒细胞介导的损伤和中性粒细胞弹性酶诱导的损伤分别有63%和64%的抑制作用。然而，50%的血清、50%的血浆或纯化的α均不能显著抑制中性粒细胞介导的损伤。1蛋白酶抑制剂。这些结果表明，在这个系统中，趋化因子刺激的人微血管内皮细胞的中性粒细胞损伤被少量LPS增强，并且可能在很大程度上是由中性粒细胞弹性酶的作用介导的。
The neutrophil has been implicated as an important mediator of vascular injury, especially after endotoxemia. This study examines neutrophil-mediated injury to human microvascular endothelial cells in vitro. We found that neutrophils stimulated by formyl-methionyl-leucyl-phenylalanine (FMLP), the complement fragment C5a, or lipopolysaccharide (LPS) (1-1,000 ng/ml) alone produced minimal endothelial injury over a 4-h assay. In contrast, neutrophils incubated with endothelial cells in the presence of low concentrations of LPS (1-10 ng/ml) could then be stimulated by FMLP or C5a to produce marked endothelial injury. Injury was maximal at concentrations of 100 ng/ml LPS and 10-7 M FMLP. Pretreatment of neutrophils with LPS resulted in a similar degree of injury, suggesting that LPS effects were largely on the neutrophil. Endothelial cell injury produced by LPS-exposed, FMLP-stimulated neutrophils had a time course similar to that induced by the addition of purified human neutrophil elastase, and different from that induced by hydrogen peroxide (H2O2). Further, neutrophil-mediated injury was not inhibited by scavengers of a variety of oxygen radical species, and occurred with neutrophils from a patient with chronic granulomatous disease, which produced no H2O2. In contrast, the specific serine elastase inhibitor methoxy-succinyl-alanyl-alanyl-prolyl-valyl-chloromethyl ketone inhibited 63% of the neutrophil-mediated injury and 64% of the neutrophil elastase-induced injury. However, neutrophil-mediated injury was not inhibited significantly by 50% serum, 50% plasma, or purified .alpha.1 proteinase inhibitor. These results suggest that, in this system, chemotactic factor-stimulated human neutrophil injury of microvascular endothelial cells is enhanced by small amounts of LPS and may be mediated in large part by the action of neutrophil elastase.