ELECTRICAL METHOD FOR DETECTION OF ENDOTHELIAL-CELL SHAPE CHANGE IN REAL-TIME - ASSESSMENT OF ENDOTHELIAL BARRIER FUNCTION
ELECTRICAL METHOD FOR DETECTION OF ENDOTHELIAL-CELL SHAPE CHANGE IN REAL-TIME - ASSESSMENT OF ENDOTHELIAL BARRIER FUNCTION
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DOI：
10.1073/pnas.89.17.7919
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发表时间：
1992-09-01
影响因子：
11.1
通讯作者：
GIAEVER, I
中科院分区：
文献类型：
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作者：
TIRUPPATHI, C;MALIK, AB;GIAEVER, I
我们已经开发了一种电学方法来研究内皮细胞形状的变化，在真实的时间，以检查在内皮屏障功能的改变的机制。通过使用在小（10（-3）cm 2）蒸发的金电极上生长的单层内皮细胞并测量电阻抗的变化来定量内皮形状变化。采用牛肺微血管内皮细胞和牛肺动脉内皮细胞，通过阻抗法研究凝血酶对细胞形态动力学的影响。α-凝血酶产生了剂量依赖性的阻抗降低，发生在0.5分钟内，在两种细胞类型，指示收缩的内皮细胞和扩大的内皮间连接，因为“四舍五入”的细胞。α-凝血酶诱导的阻抗降低持续几乎等于2小时，之后该值恢复到基础水平。用蛋白激酶C抑制剂、钙磷蛋白C或8-溴腺苷3 '，5'-环一磷酸预处理内皮细胞可防止阻抗降低，这表明内皮细胞的变化是由第二信使途径的激活调节的。α-凝血酶诱导的阻抗降低与先前观察到的跨内皮白蛋白渗透性增加和α-凝血酶激发后细胞间间隙形成的证据一致。阻抗测量可能是一种有价值的体外方法，用于评估炎症介质引起的内皮屏障功能降低的机制。由于迅速发生的变化，在内皮细胞的形状，如凝血酶介导的第二信使途径的激活介质，监测内皮细胞动力学的能力，在真实的时间可以提供深入了解的信号转导事件介导的内皮通透性增加。
We have developed an electrical method to study endothelial cell shape changes in real time in order to examine the mechanisms of alterations in the endothelial barrier function. Endothelial shape changes were quantified by using a monolayer of endothelial cells grown on a small (10(-3) cm2) evaporated gold electrode and measuring the changes in electrical impedance. Bovine pulmonary microvessel endothelial cells and bovine pulmonary artery endothelial cells were used to study the effects of alpha-thrombin on cell-shape dynamics by the impedance measurement. Alpha-thrombin produced a dose-dependent decrease in impedance that occurred within 0.5 min in both cell types, indicative of retraction of endothelial cells and widening of interendothelial junctions because of "rounding up" of the cells. The alpha-thrombin-induced decrease in impedance persisted for almost-equal-to 2 hr, after which the value recovered to basal levels. Pretreatment of endothelial cells with the protein kinase C inhibitor, calphostin C, or with 8-bromoadenosine 3',5'-cyclic monophosphate prevented the decreased impedance, suggesting that the endothelial cell change is modulated by activation of second-messenger pathways. The alpha-thrombin-induced decrease in impedance was in agreement with the previously observed increases in transendothelial albumin permeability and evidence of formation of intercellular gaps after alpha-thrombin challenge. The impedance measurement may be a valuable in vitro method for the assessment of mechanisms of decreased endothelial barrier function occurring with inflammatory mediators. Since the rapidly occurring changes in endothelial cell shape in response to mediators such as thrombin are mediated activation of second-messenger pathways, the ability to monitor endothelial cell dynamics in real time may provide insights into the signal-transduction events mediating the increased endothelial permeability.