Cerini Claire, Dou Laetitia, Anfosso Francine, Sabatier Florence, Moal Valérie, Glorieux Griet, De Smet Rita, Vanholder Raymond, Dignat-George Françoise, Sampol José, Berland Yvon, Brunet Philippe
INSERM EMI0019, Faculté de Pharmacie, Université e la Méditerranée, 27 bld Jean Moulin, 13385 Marseille, Cedex 5, France.
Thromb Haemost. 2004 Jul;92(1):140-50. doi: 10.1160/TH03-07-0491.
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.
慢性肾衰竭（CRF）患者存在内皮功能障碍，这可能与血液和组织中蓄积的尿毒症潴留溶质有关。在本研究中，我们调查了尿毒症潴留溶质对甲酚在体外对内皮细胞（人脐静脉内皮细胞）屏障功能的影响。对甲酚的测试浓度为CRF患者体内的浓度，由于对甲酚与蛋白质结合，因此在有和没有生理浓度人白蛋白（4 g/dl）的情况下进行实验。加入白蛋白后，我们发现对甲酚在暴露24小时后会导致内皮通透性大幅增加。伴随着内皮通透性的增加，对甲酚诱导肌动蛋白细胞骨架重排以及黏附连接改变。这些分子事件表现为皮质肌动蛋白染色减少，与细胞内应力纤维的形成有关，以及连接蛋白血管内皮钙黏蛋白染色减少。连接蛋白血管内皮钙黏蛋白染色减少与膜表达降低无关。不加入白蛋白时，对甲酚的作用更明显。特异性Rho激酶抑制剂Y-27632可抑制对甲酚的作用，表明对甲酚以Rho激酶依赖的方式介导内皮通透性增加。总之，这些结果表明对甲酚在体外可导致内皮屏障功能严重障碍，并提示这种尿毒症潴留溶质可能参与了CRF患者中观察到的内皮功能障碍。
