Effect of vascular endothelial growth factor on cultured endothelial cell monolayer transport properties
Effect of vascular endothelial growth factor on cultured endothelial cell monolayer transport properties
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DOI：
10.1006/mvre.1999.2225
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发表时间：
2000-03-01
影响因子：
3.1
通讯作者：
Tarbell, JM
中科院分区：
文献类型：
--
作者：
Chang, YS;Munn, LL;Tarbell, JM
血管内皮生长因子(VEGF)是体内有效的微血管通透性增强剂。到目前为止，它对血管内皮细胞单层的水力传导性(L-p)和扩散白蛋白通透性(P-e)的影响还没有得到全面的体外评估。我们假设，根据血管位置和内皮表型的不同，血管内皮生长因子对内皮运输特性的影响也不同。使用三种成熟的内皮细胞培养模型-人脐静脉内皮细胞(HUVECs)、牛主动脉内皮细胞(BAECs)和牛视网膜微血管细胞(BRECs)-生长在多孔聚碳酸酯过滤器上，我们能够产生具有限制性屏障特征的基线传输特性。我们的结果显示，在血管内皮细胞生长因子(100 ng/ml)作用3h后，内皮细胞内皮细胞L-p的表达分别增加3.1倍和5.7倍。而血管内皮细胞与血管内皮细胞共同孵育3h(10 0 ng/ml)或2 4h(2 5 ng/ml)后，即使对炎症介质、凝血酶(1U/ml，L-p在2 5min内增加2 7倍)和缓激肽(10 mU/m l，L-p在2 0min内增加4倍)也有反应，但在3h(10 0 ng/ml)或2 4h(2 5 ng/ml)时，细胞内皮细胞的L-p水平无明显变化。蛋白激酶C(PKC)和一氧化氮(NO)是血管内皮生长因子信号转导的下游效应分子。BAEC L-p对NO(SNAP)和PKC(PMA)的激活有反应，而对HWEC L-p无影响。此外，蛋白激酶C抑制剂星形孢菌素(50 ng/ml)可显著抑制血管内皮生长因子诱导的L-β细胞的增殖，而用一氧化氮合酶抑制剂L(100mU M)抑制一氧化氮合酶对血管内皮细胞生长因子诱导的L-β细胞增殖无影响。这些数据提供了强有力的证据表明，血管内皮细胞中血管内皮生长因子诱导的L-p的增加是由蛋白激酶C依赖的机制介导的。在弥漫性白蛋白P-e方面，3h时BAECs和BRECs的P-e分别增加了6.0倍和9.9倍，而在3h(100 ng/ml)和24 h(25 ng/ml)时，VEGF对P-e的影响不明显。综上所述，这些数据表明，根据血管类型的不同，血管内皮生长因子对内皮运输特性的影响是不同的，细胞信号通路的不同是血管内皮生长因子反应性差异的基础。(C)2000年学术出版社。
Vascular endothelial growth factor (VEGF) is a potent enhancer of microvascular permeability in vivo. To date, its effects on hydraulic conductivity (L-p) and diffusive albumin permeability (P-e) of endothelial monolayers have not been thoroughly assessed in vitro. We hypothesized that VEGF affects endothelial transport properties differently depending on vessel location and endothelial phenotype. Using three well-established endothelial cell culture models-human umbilical vein endothelial cells (HUVECs), bovine aortic endothelial cells (BAECs), and bovine retinal microvascular cells (BRECs)-grown on porous, polycarbonate filters we were able to produce baseline transport properties characteristic of restrictive barriers. Our results show 3.1-fold and 5.7-fold increases in endothelial L-p for BAEC and BREC monolayers, respectively, at the end of 3 h of VEGF (100 ng/ml) exposure. HUVECs, however, showed no significant alteration in L-p after 3 h (100 ng/ml) or 24 h (25 ng/ml) of incubation with VEGF even though they were responsive to the inflammatory mediators, thrombin (1 U/ml; 27-fold increase in L-p in 25 min) and bradykinin (10 mu M; 4-fold increase in L-p in 20 min). Protein kinase C (PKC) and nitric oxide (NO) are downstream effecters of VEGF signaling. BAEC L-p was responsive to activation of NO (SNAP) and PKC (PMA), whereas these agents had no effect in altering HWEC L-p. Moreover, BAECs exposed to the PKC inhibitor, staurosporine (50 ng/ml), exhibited significant attenuation of VEGF-induced increase in L-p, but inhibition of nitric oxide synthase (NOS) with L-NMMA (100 mu M) had no effect in altering the VEGF-induced increase in L-p. These data provide strong evidence that in BAECs, the VEGF-induced increase in L-p is mediated by a PKC-dependent mechanism. Regarding diffusive albumin P-e, at the end of 3 h, BAECs and BRECs showed 6.0-fold and 9.9-fold increases in P-e in response to VEGF (100 ng/ml), whereas VEGF had no significant effect after 3 h (100 ng/ml) or 24 h (25 ng/ml) in changing HWEC P-e. In summary, these data indicate that VEGF affects endothelial transport properties differently depending on the vessel type and that differences in cell signaling pathways underlie the differences in VEGF responsiveness. (C) 2000 Academic Press.