TNF-alpha potentiates oxidant and reperfusion-induced endothelial cell injury
TNF-alpha potentiates oxidant and reperfusion-induced endothelial cell injury
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DOI：
10.1006/jsre.1996.0101
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发表时间：
1996-02-15
影响因子：
2.2
通讯作者：
Rees, RS
中科院分区：
文献类型：
--
作者：
Gilmont, RR;Dardano, A;Rees, RS
再灌注后的肺水肿是一个主要的临床问题。氧化损伤引起的内皮细胞形状的变化可能是与再灌注损伤相关的直接毛细血管渗漏的原因。在这些实验中，我们研究了肿瘤坏死因子-α (TNF-α) 在缺血再灌注后急性内皮细胞损伤中的作用。在产生远端缺血之前，用针对 TNF-α 的中和抗血清处理 Sprague-Dawley 大鼠。与接受相同程序但未进行抗血清治疗的大鼠相比，这些大鼠在 4 小时缺血和 30 分钟再灌注后的急性肺水肿显着减轻（P < 0.05）。开发了体外模型以确定 TNF-α 是否对内皮细胞对缺血再灌注的反应有直接影响。测量了 TNF-α 和氧化应激对培养的内皮细胞单层完整性的影响。大鼠肺动脉内皮细胞单层在体外通过增加通透性对氧化应激做出反应。当这些细胞暴露于 50 μM 过氧化氢 (H2O2) 或来自 Sprague-Dawley 大鼠缺血后肢的血浆 (50 μl/ml) 时，细胞形状发生改变，并且 I-125-白蛋白在细胞单层中的扩散增加。用低水平的重组小鼠TNF-α预处理培养的细胞显着影响细胞形状的变化和通透性的增加（P < 0.05）。根据培养基乳酸脱氢酶水平确定，体外细胞单层的通透性增加并非由于细胞裂解所致。这种效应似乎是由于使用视频延时显微镜观察到的细胞变圆和收缩所致。这些数据表明TNF-α对内皮细胞有直接作用，从而使细胞更容易受到再灌注伴随的氧化损伤。 (C) 1996 学术出版社
Pulmonary edema following reperfusion is a major clinical problem. Changes in endothelial cell shape induced by oxidant injury may account for immediate capillary leakage associated with reperfusion injury. In these experiments we examined the role of tumor necrosis factor-alpha (TNF-alpha) in acute endothelial cell injury following ischemia-reperfusion. Sprague-Dawley rats were treated with a neutralizing antisera directed against TNF-alpha prior to production of distal ischemia. These rats demonstrated a significant reduction (P < 0.05) in acute lung edema in response to 4 hr of ischemia and 30 min of reperfusion when compared to rats undergoing the same procedure without antisera treatment. An in vitro model was developed to determine if TNF-alpha had a direct effect on endothelial cell response to ischemia-reperfusion. The effects of TNF-alpha and oxidant stress on the integrity of cultured endothelial cell monolayers was measured. Rat pulmonary artery endothelial cell monolayers reacted in vitro to oxidant stress by an increase in permeability. The cells changed shape and an increase in diffusion of I-125-albumin across cell monolayers resulted when these cells were exposed to 50 mu M hydrogen peroxide (H2O2) or plasma from the ischemic hind limb of a Sprague-Dawley rat (50 mu l/ml). Pretreatment of cultured cells with low levels of recombinant mouse TNF-alpha significantly affected both the cell shape change and the increase in permeability (P < 0.05). Increased permeability of cell monolayers in vitro was not due to cell lysis as determined by media lactate dehydrogenase levels. The effect appeared to be due to cellular rounding and contraction seen using video time lapse microscopy. These data suggest a direct effect of TNF-alpha on endothelial cells, whereby the cells are rendered more susceptible to oxidant injury accompanying reperfusion. (C) 1996 Academic Press,Inc.