P-cresol, a uremic retention solute, alters the endothelial barrier function in vitro
P-cresol, a uremic retention solute, alters the endothelial barrier function in vitro
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DOI：
10.1160/th03-07-0491
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发表时间：
2004-07-01
影响因子：
6.7
通讯作者：
Brunet, P
中科院分区：
文献类型：
--
作者：
Cerini, C;Dou, L;Brunet, P
慢性肾衰竭（CRF）患者表现出内皮功能障碍，这可能涉及在血液和组织中蓄积的尿毒症保留溶质。在这项研究中，我们研究了尿毒症滞留溶质对甲酚对内皮细胞（HUVEC）屏障功能的体外影响。在CRF患者中发现的浓度下测试对甲酚，并且由于对甲酚是蛋白质结合的，因此在具有和不具有生理浓度的人白蛋白（4g/dl）的情况下进行实验。使用白蛋白时，我们发现对甲酚暴露24小时后会导致内皮通透性大幅增加。伴随着这种内皮通透性的增加，对甲酚诱导肌动蛋白细胞骨架的重组和粘附连接的改变。这些分子的事件证明了减少染色的皮质肌动蛋白，与整个细胞的应力纤维的形成，并通过减少染色的连接VE-钙粘蛋白。连接VE-钙粘蛋白染色的减少与膜表达的减少无关。在不含白蛋白的情况下，对甲酚的作用更为明显。特异性Rho激酶抑制剂Y-27632抑制对甲酚的作用，表明对甲酚以Rho激酶依赖性方式介导内皮通透性的增加。总之，这些结果表明，对甲酚在体外引起内皮屏障功能的严重功能障碍，并表明这种尿毒症滞留溶质可能参与CRF患者中观察到的内皮功能障碍。
Patients with chronic renal failure (CRF) exhibit endothelial dysfunction, which may involve uremic retention solutes that accumulate in blood and tissues. In this study, we investigated the in vitro effect of the uremic retention solute p-cresol on the barrier function of endothelial cells (HUVEC). P-cresol was tested at concentrations found in CRF patients, and since p-cresol is protein-bound, experiments were performed with and Without physiological concentration of human albumin (4 g/dl). With albumin, we showed that p-cresol caused a strong increase in endothelial permeability after a 24-hour exposure. Concomitant with this increase in endothelial permeability, p-cresol induced a reorganization of the actin cytoskeleton and an alteration of adherens junctions. These molecular events were demonstrated by the decreased staining of cortical actin, associated with the formation of stress fibers across the cell, and by the decreased staining of junctional VE-cadherin. This decrease in junctional VE-cadherin staining was not associated with a reduction of membrane expression. Without albumin, the effects of p-cresol were more pronounced. The specific Rho kinase inhibitor, Y-27632, inhibited the effects of p-cresol, indicating that p-cresol mediates the increase in endothelial permeability in a Rho kinase-dependent way. In conclusion, these results show that p-cresol causes a severe dysfunction of endothelial barrier function in vitro and suggest this uremic retention solute may participate in the endothelium dysfunction observed in CRF patients.