Endothelial junctional protein redistribution and increased monolayer permeability in human umbilical vein endothelial cells isolated during preeclampsia
Endothelial junctional protein redistribution and increased monolayer permeability in human umbilical vein endothelial cells isolated during preeclampsia
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DOI：
10.1067/mob.2002.119638
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发表时间：
2002-02-01
影响因子：
9.8
通讯作者：
Alexander, JS
中科院分区：
文献类型：
--
作者：
Wang, YP;Gu, Y;Alexander, JS
目的：检测从先兆子痫妇女和正常妊娠妇女分离的内皮细胞单层通透性和连接蛋白的分布和表达。我们假设，增加内皮细胞单层渗透性先兆子痫反映了改变单层屏障性能所产生的解体的内皮细胞连接proteins.Study设计：内皮细胞分离脐静脉正常妊娠妇女（n = 9）和妇女先兆子痫（n = 9）后立即交付。在内皮细胞的第一次传代中，通过测量辣根过氧化物酶通过在transwell滤器上生长的汇合细胞单层来确定渗透性，并通过使用蛋白质的免疫荧光染色和Western印迹分析来评估血管内皮钙粘蛋白和闭合蛋白的分布和蛋白表达。血管内皮钙粘蛋白的分布也进行了评估，在第二和第三代内皮细胞。逆转录聚合酶链反应检测血管内皮钙粘蛋白和闭合蛋白的信使核糖核酸表达。结果：与正常妊娠妇女相比，先兆子痫妇女内皮细胞单层相对通透性显著增加（DeltaOD（470）为0.271 ± 0.049 vs 0.086 ± 0.031; P
OBJECTIVE: To examine monolayer permeability and junctional protein distribution and expression in endothelial cells isolated from women with preeclampsia and from women with normal pregnancies. We hypothesized that increased endothelial monolayer permeability in preeclampsia reflects altered monolayer barrier properties produced by disorganization of endothelial cell junction proteins.STUDY DESIGN: Endothelial cells were isolated from umbilical veins from women with normal pregnancies (n = 9) and from women with preeclampsia (n = 9) immediately after delivery. In the first passage of endothelial cells, the permeability was determined by measurement of horseradish peroxidase passage through confluent cell monolayers grown on transwell filters, and the distribution and protein expression of vascular endothelial cadherin and occludin were evaluated by use of immunofluorescent staining of the proteins and Western blot analysis. The distribution of vascular endothelial cadherin was also evaluated in the second and third passage endothelial cells. Messenger ribonucleic acid expression of vascular endothelial cadherin and occludin were examined by reverse transcriptase polymerase chain reaction. Data were expressed as mean values ( SE) and were analyzed by use of an unpaired t test.RESULTS: The relative monolayer permeability was significantly increased in the endothelial cells isolated from women with preeclampsia compared with those isolated from women with normal pregnancies (0.271 +/- 0.049 versus 0.086 +/- 0.031 for DeltaOD(470); P