Institute of Biomedical Science, Hanyang University College of Medicine, Seoul, Korea.
Department of Internal Medicine, Hanyang University College of Medicine, Seoul, Korea.
Ren Fail. 2019 Nov;41(1):1011-1015. doi: 10.1080/0886022X.2019.1686018. Epub 2019 Nov 14.
We hypothesized that tight junction (TJ) proteins may have a role in paracellular transport of solute and water in peritoneal dialysis (PD) patients. Previous studies on TJ proteins in PD patients have used only cultured human peritoneal mesothelial cells (HPMCs). This study was undertaken to test whether TJ proteins are directly identified from PD effluent and whether their expressions are associated with functional parameters of PD. Dialysis effluents were collected from 40 patients undergoing PD, after the peritoneal equilibration test (PET). Different molecular sizes of Amicon Ultra-15 Centrifugal Filter Units were used to concentrate and purify proteins in PD effluents, and immunoblot analyses for occludin, ZO-1, and claudins were carried out to test for their existence and relationships with peritoneal clearance or results of the PET. Immunoblotting from PD effluents revealed discrete bands of occludin (∼65 kDa), ZO-1 (∼215 kDa), claudin-1 (∼22 kDa), and claudin-15 (∼22 kDa) in all 40 patients. The peritoneal creatinine clearance inversely correlated with the protein expression of claudin-1 (= -0.369, = .019), and the dialysate-to-plasma creatinine ratio at 4 h PET correlated with occludin ( = 0.396, = .011) and inversely correlated with claudin-15 (= -0.393, = .012). In PD patients, expression of peritoneal TJ proteins can be estimated from the dialysis effluent and may be used as novel peritoneal biomarkers.
我们假设紧密连接 (TJ) 蛋白在腹膜透析 (PD) 患者的溶质和水的旁细胞转运中可能具有作用。以前对 PD 患者 TJ 蛋白的研究仅使用培养的人腹膜间皮细胞 (HPMCs)。本研究旨在测试 TJ 蛋白是否可直接从 PD 流出物中鉴定出来,以及它们的表达是否与 PD 的功能参数相关。在腹膜平衡试验 (PET) 后,从 40 名接受 PD 的患者中收集透析流出物。使用不同分子量的 Amicon Ultra-15 离心超滤单元浓缩和纯化 PD 流出物中的蛋白质,并进行免疫印迹分析以检测 occludin、ZO-1 和 claudins 的存在及其与腹膜清除率或 PET 结果的关系。从 PD 流出物的免疫印迹显示,在所有 40 名患者中均存在离散的 occludin(∼65 kDa)、ZO-1(∼215 kDa)、claudin-1(∼22 kDa)和 claudin-15(∼22 kDa)条带。腹膜肌酐清除率与 claudin-1 的蛋白表达呈负相关(= -0.369,=.019),4 h PET 时的透析液-血浆肌酐比值与 occludin 呈正相关(= 0.396,=.011),与 claudin-15 呈负相关(= -0.393,=.012)。在 PD 患者中,可从透析流出物中估计腹膜 TJ 蛋白的表达,并可作为新型腹膜生物标志物。