TNF-ALPHA INDUCES ENDOTHELIAL-CELL F-ACTIN DEPOLYMERIZATION, NEW ACTIN SYNTHESIS, AND BARRIER DYSFUNCTION
TNF-ALPHA INDUCES ENDOTHELIAL-CELL F-ACTIN DEPOLYMERIZATION, NEW ACTIN SYNTHESIS, AND BARRIER DYSFUNCTION
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DOI：
10.1152/ajpcell.1993.264.4.c894
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发表时间：
1993-04-01
影响因子：
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通讯作者：
CAMPBELLWASHINGTON, J
中科院分区：
文献类型：
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作者：
GOLDBLUM, SE;DING, XD;CAMPBELLWASHINGTON, J
肿瘤坏死因子-α在体外影响肺血管内皮细胞屏障功能。我们研究了重组的肿瘤坏死因子-α(rTNF-α)是否通过肌动蛋白重组来调节内皮屏障功能。将融合后的牛肺动脉内皮细胞单层暴露于人重组肿瘤坏死因子-α(1000U/ml)中，观察1)跨内皮[C-14]白蛋白流量，2)荧光显微镜下F-肌动蛋白的形成，3)荧光分光光度法定量F-肌动蛋白，4)脱氧核糖核酸酶I抑制法检测单体G-肌动蛋白水平。在此期间，内皮F-肌动蛋白池减少(P=0.0064)，G-肌动蛋白池相对增加(P<；0.0001)。先前用鬼臼毒素稳定F-肌动蛋白可防止重组肿瘤坏死因子-α诱导的G-肌动蛋白增加(P<；0.002)以及屏障功能的改变(P<；0.01)。预先抑制蛋白质合成增强了重组肿瘤坏死因子-α诱导的F-肌动蛋白的减少(P<；0.0001)，钝化了G-肌动蛋白的增加(P<；0.002)，并增加了重组肿瘤坏死因子-α诱导的内皮屏障功能的改变(P<；0.003)。因此，rTNF-α诱导肺血管内皮细胞F-肌动蛋白解聚、细胞间隙形成和屏障功能障碍。重组肿瘤坏死因子-α还增加了总肌动蛋白(P<；0.02)和新肌动蛋白合成(P<；0.002)，这可能是内皮细胞对重组肿瘤坏死因子-α诱导的F-肌动蛋白解聚的代偿反应。
Tumor necrosis factor-alpha (TNF-alpha) influences pulmonary vascular endothelial barrier function in vitro. We studied whether recombinant TNF-alpha (rTNF-alpha) regulates endothelial barrier function through actin reorganization. Postconfluent bovine pulmonary artery endothelial cell monolayers were exposed to human rTNF-alpha (1,000 U/ml) and evaluated for 1) transendothelial [C-14]albumin flux, 2) F-actin organization with fluorescence microscopy, 3) F-actin quantitation by spectrofluorometry, and 4) monomeric G-actin levels by the deoxyribonuclease I inhibition assay. rTNF-alpha induced increments in [C-14]albumin flux (P < 0.04) and intercellular gap formation at greater-than-or-equal-to 2-6 h. During this same time, the endothelial F-actin pool decreased (P = 0.0064), with reciprocal increases in the G-actin pool (P < 0.0001). Prior F-actin stabilization with phallicidin protected against the rTNF-alpha-induced increments in G-actin (P < 0.002) as well as changes in barrier function (P < 0.01). Prior protein synthesis inhibition enhanced the rTNF-alpha-induced decrement in F-actin (P < 0.0001), blunted the G-actin increment (P < 0.002), and increased rTNF-alpha-induced changes in endothelial barrier function (P < 0.003). Therefore, rTNF-alpha induces pulmonary vascular endothelial F-actin depolymerization, intercellular gap formation, and barrier dysfunction. rTNF-alpha also increased total actin (P < 0.02) and new actin synthesis (P < 0.002), which may be a compensatory endothelial cell response to rTNF-alpha-induced F-actin depolymerization.