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首页> 外文期刊>Journal of Cellular Physiology >Mechanisms of alpha-thrombin, histamine, and bradykinin induced endothelial permeability.
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Mechanisms of alpha-thrombin, histamine, and bradykinin induced endothelial permeability.

机译：alpha-thrombin机制,组胺缓激肽诱导内皮渗透性。

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alpha-Thrombin, bradykinin, and histamine are endogenous mediators that increase endothelial permeability. We examined the mechanism by which these three vasoactive mediators could alter permeability to albumin of human umbilical vein endothelial cells (HUVEC). HUVEC were grown to confluence on Transwell membranes and we monitored the flux of fluorescein isothiocyanate-labeled human serum albumin across the membrane from the upper to lower chamber of the Transwell. Addition of alpha-thrombin, bradykinin, or histamine increased the permeability coefficient of the HUVEC monolayer. At 30 min the permeability coefficient for alpha-thrombin was 4.92 x 10(-6) cm/sec while histamine was 4.47 x 10(-6) cm/sec. Maximum changes in the permeability coefficient were about three-fold control baseline values (1.59 x 10(-6) cm/sec). There was also a temporal difference in the magnitude of the permeability coefficient. alpha-Thrombin and bradykinin induced HUVEC permeability was increased for the first 90 min after which it returned to control levels. In contrast, histamine increased the permeability of the HUVEC monolayer throughout the 2 h experiment. To determine a possible intracellular mechanism of the altered permeability coefficients, HUVEC were labeled with FURA-2 and intracellular calcium was monitored by digital fluorescence ratio imaging. Maximum intracellular calcium in HUVEC was increased by alpha-thrombin (245 +/- 20 nM) and histamine (210 +/- 22 nM), but not by bradykinin (70 +/- 7 nM) as compared to control (69 +/- 10). Fluorescent photomicrographs of HUVEC stimulated with the three agonists indicated that alpha-thrombin and histamine substantially altered HUVEC f-actin arrangement, while bradykinin had no effect on HUVEC f-actin distribution. These data support previous in vitro and in vivo studies demonstrating increased permeability by all three agonists. These data also show, for the first time, that histamine and alpha-thrombin increased permeability by calcium-dependent intracellular pathways, but bradykinin operates through a calcium-independent mechanism.
机译：alpha-Thrombin、缓激肽、组胺内源性介质,增加内皮渗透率。这三种血管活性的介质可能会改变渗透到人类脐静脉的白蛋白血管内皮细胞(HUVEC)。融合Transwell膜和我们检测荧光素的通量isothiocyanate-labeled人类血清白蛋白细胞膜上降低商会Transwell。缓激肽、组胺的增加渗透系数的HUVEC单层。在30分钟的渗透系数alpha-thrombin 4.92 x 10(6)厘米/秒组胺是4.47 x 10(6)厘米/秒。渗透系数的变化三倍(1.59 x控制基线值10(6)厘米/秒)。不同渗透率的大小系数。诱导HUVEC渗透率的增加前90分钟后返回控制的水平。HUVEC的渗透率单层2 h的实验。细胞内改变的机制渗透系数,HUVEC标记FURA-2和细胞内钙通过数字荧光比率成像监测。最大的在HUVEC细胞内钙增加了alpha-thrombin(245 + / - 20海里)组胺(210 + / - 22 nM),但不是由缓激肽(70 + / - 7海里)与控制(69 + / - 10)。荧光显微照片HUVEC的刺激三个主人公表示alpha-thrombin和组胺显著改变HUVEC f -肌动蛋白安排,缓激肽对HUVEC f -肌动蛋白没有影响分布。体外和体内研究证明增加通过这三种受体激动剂渗透性。第一次还显示,组胺和alpha-thrombin渗透率增加了calcium-dependent细胞内途径,但是缓激肽通过calcium-independent运作机制。

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