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首页> 外文期刊>American Journal of Physiology >Thrombin enhances the barrier function of rat microvascular endothelium in a PAR~(-1)-dependent manner
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Thrombin enhances the barrier function of rat microvascular endothelium in a PAR~(-1)-dependent manner

机译：凝血酶在PAR〜（-1）依赖性的情况下提高大鼠微血管内皮的阻隔功能

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First published December 14, 2007; doi:10.1152/ajplung.00107.2007.-Thrombin is a multifunctional coagulation protease with pro- and anti-inflammatory vascular effects. We questioned whether thrombin may have segmentally differentiated effects on pulmonary endothelium. In cultured rat endothelial cells, rat thrombin (10 U/ml) recapitulated the previously reported decrease in transmonolayer electrical resistance (TER), F-actin stress fiber formation, paracellular gap formation, and increased permeability. In contrast, in rat pulmonary microvascular endothelial cells (PMVEC), isolated on the basis of Griffonia simplicifolia lectin recognition, thrombin increased TER, induced fewer stress fibers, and decreased permeability. To assess for differential proteinase-activated receptor (PAR) expression as a basis for the different responses, PAR family expression was analyzed. Both pulmonary artery endothelial cells and PMVEC expressed PAR~(-1) and PAR-2; however, only PMVEC expressed PAR-3, as shown by both RT-PCR and Western analysis. PAR~(-1) activating peptides (PAR-APs: SFLLRN-NH2 and TFLLRN-NH2) were used to confirm a role for the PAR~(-1) receptor. PAR-APs (25-250 jiM) also increased TER, formed fewer stress fibers, and did not induce paracellular gaps in PMVEC in contrast to that shown in pulmonary artery endothelial cells. These results were confirmed in isolated perfused rat lung preparations. PAR-APs (100 muml) induced a 60% increase in the filtration coefficient over baseline. However, by transmission electron microscopy, perivascular fluid cuffs were seen only along conduit veins and arteries without evidence of intra-alveolar edema. We conclude that thrombin exerts a segmentally differentiated effect on endothelial barrier function in'vitro, which corresponds to a pattern of predominant perivascular - fluid cuff formation in situ. This may indicate a distinct role for thrombin in the microcirculation.
机译：2007年12月14日第一次出版; DOI：10.1152 / AJPLUNG.00107.2007.-凝血酶是一种多功能凝血蛋白酶，具有促炎和抗炎血管作用。我们质疑凝血酶是否可能对肺内皮细胞分段分化的影响。在培养的大鼠内皮细胞中，大鼠凝血酶（10U / mL）重新携带先前报道的透射糖层电阻（TER）的降低，F-actin应力纤维形成，细胞间隙形成和渗透性增加。相比之下，在大鼠肺部微血管内皮细胞（PMVEC）中，在Griffinia Simplecolia凝集素识别的基础上分离，凝血酶增加，诱导较少的应力纤维，并降低渗透性。评估差异蛋白酶活化受体（PAR）表达作为不同反应的基础，分析了PAR系列表达。肺动脉内皮细胞和PMVEC都表达了PAR〜（-1）和PAR-2;然而，只有PMVEC表达PAR-3，如RT-PCR和西方分析所示。 PAR〜（-1）活化肽（PAR-AP：SFLLRN-NH2和TFLLRRN-NH2）用于证实PAR〜（-1）受体的作用。 PAR-AP（25-250吉姆）也增加了三倍，形成了较少的应力纤维，并且与肺动脉内皮细胞中所示的相反，没有诱导PMVEC中的肺膜间隙。在分离的灌注大鼠肺制剂中证实了这些结果。 PAR-AP（100 muml）诱导基线过滤系数增加60％。然而，通过透射电子显微镜，仅沿着导管静脉和动脉观察血管液袖口，没有肺泡内水肿的证据。我们得出结论，凝血酶对血管内屏障函数的分段分化作用，其对应于原位的主要血管 - 流体袖带的模式。这可能表明微循环中的凝血酶具有不同的作用。

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