Idell S, Gonzalez K K, MacArthur C K, Gillies C, Walsh P N, McLarty J, Thrall R S
Am Rev Respir Dis. 1987 Jul;136(1):124-33. doi: 10.1164/ajrccm/136.1.124.
Although pulmonary fibrin deposition and coagulation abnormalities have been observed in acute lung injury in humans, their role in the pathogenesis of pulmonary disorders is unclear. In order to gain further insights into the role of the coagulation in lung injury, we examined the relationship between procoagulant activity in bronchoalveolar lavage (BAL) fluids and the evolution of bleomycin-induced lung injury in marmosets. The BAL procoagulant activity was increased at 1, 2, and 4 wk after bleomycin challenge compared with that in control subjects, and it was capable of shortening the recalcification times of plasmas deficient in factor VII and factor VIII but not in factor X. This profile suggested the presence in BAL of an activator of factor X. Activation of purified human factor X by BAL was demonstrated by measuring the amidolytic activity of the generated factor Xa on its N-benzoyl-L-isoleucyl L-glutamyl-glycyl-L-argenine-p-nitroanilide substrate. Factor X activating activity was increased in BAL at 2 wk after bleomycin challenge. Cleavage of 125I-labeled human factor X by BAL from bleomycin-challenged marmosets yielded a 55,500 Mr product that comigrated with factor Xa, the appearance of which correlated strongly with amidolytic evidence of factor Xa activity. Electron microscopy of the lungs of animals from all groups revealed pulmonary fibrin deposition at 2 wk after bleomycin challenge, at the time of increased BAL procoagulant and factor X activating activity. The BAL procoagulant activity was completely sedimentable by ultracentrifugation and was inhibited by concanavalin A and phospholipase C. Activation of purified factor X by BAL was inhibited by monospecific polyclonal goat and rabbit antibodies to human factor VII as well as antibody to bovine tissue factor, demonstrating that factor X activating activity in BAL was attributable to tissue factor associated with material similar to factors VII or VIIa. We conclude that procoagulant activity in BAL increases after bleomycin challenge in marmosets and is attributable to activation of factor X by tissue factor associated with factors VII or VIIa-like material. Increased BAL procoagulant activity is temporally associated with pulmonary fibrin deposition and pulmonary fibrosis during bleomycin-induced pulmonary injury in the marmoset.
尽管在人类急性肺损伤中已观察到肺内纤维蛋白沉积和凝血异常，但其在肺部疾病发病机制中的作用尚不清楚。为了进一步深入了解凝血在肺损伤中的作用，我们研究了支气管肺泡灌洗（BAL）液中的促凝活性与博来霉素诱导的狨猴肺损伤进展之间的关系。与对照组相比，博来霉素攻击后1周、2周和4周时BAL促凝活性增加，且其能够缩短缺乏因子VII和因子VIII但不缺乏因子X的血浆的复钙时间。这一特征提示BAL中存在因子X激活剂。通过测量生成的因子Xa对其N-苯甲酰-L-异亮氨酰-L-谷氨酰-甘氨酰-L-精氨酸-对硝基苯胺底物的酰胺水解活性，证实了BAL对纯化的人因子X的激活作用。博来霉素攻击后2周时BAL中的因子X激活活性增加。来自博来霉素攻击的狨猴的BAL对125I标记的人因子X的切割产生了一个55,500 Mr的产物，其迁移率与因子Xa相同，其出现与因子Xa活性的酰胺水解证据密切相关。对所有组动物肺部的电子显微镜检查显示，在博来霉素攻击后2周，即BAL促凝活性和因子X激活活性增加时，出现了肺内纤维蛋白沉积。BAL促凝活性可通过超速离心完全沉淀，并被伴刀豆球蛋白A和磷脂酶C抑制。BAL对纯化因子X的激活被针对人因子VII的单特异性多克隆山羊和兔抗体以及针对牛组织因子的抗体所抑制，表明BAL中的因子X激活活性归因于与类似于因子VII或VIIa的物质相关的组织因子。我们得出结论，博来霉素攻击后狨猴BAL中的促凝活性增加，且归因于与因子VII或VIIa样物质相关的组织因子对因子X的激活。在博来霉素诱导的狨猴肺损伤期间，BAL促凝活性增加与肺内纤维蛋白沉积和肺纤维化在时间上相关。
