Atmani F, Glenton P A, Khan S R
Department of Pathology, College of Medicine, University of Florida, Gainesville 32610-0275, USA.
Urol Res. 1998;26(3):201-7. doi: 10.1007/s002400050047.
The purpose of our study was to identify the proteins and investigate the differences, if any, between protein components of the matrices of calcium oxalate (CaOx) and calcium phosphate (CaP) crystals induced in vitro in whole human urine of healthy individuals and kidney stone patients. In addition, preliminary studies were performed to understand the effect of centrifugation and filtration of urine on its protein contents. Crystallization in urine was induced by addition of an oxalate or phosphate load. Crystals were collected, washed, and analyzed by scanning electron microscopy, X-ray diffraction, and energy dispersive X-ray microanalysis. Matrix proteins were obtained by demineralization with ethylene diamine tetraacetic acid (EDTA), analyzed by polyacrylamide gel electrophoresis, and identified by western blotting technique. No significant differences were detected between protein components of the matrices of CaOx and CaP crystals and between the crystal matrices obtained from the urine of normal and stone forming subjects. Albumin (AB), inter-alpha-inhibitor (IalphaI) related proteins, alpha-1 microglobulin (alpha-1 m), osteopontin (OPN), prothrombin (PT)-related proteins and Tamm-Horsfall protein (THP) were identified in matrices of both CaOx and CaP crystals induced in urine from both the normal subjects and stone formers. AB, PT-related proteins and OPN were the main constituents. The other proteins were present in smaller but detectable amounts. However, CaP crystal matrix, contained a large amount of THP. In addition CaP crystals contained significantly more proteins than CaOx crystals. Centrifugation and/or filtration of the urine resulted in reduction of many high molecular weight proteins including THP, AB and OPN in the urine.
我们研究的目的是鉴定蛋白质，并调查在健康个体和肾结石患者的全人类尿液中体外诱导生成的草酸钙（CaOx）和磷酸钙（CaP）晶体基质的蛋白质成分之间是否存在差异（若有差异）。此外，还进行了初步研究以了解尿液离心和过滤对其蛋白质含量的影响。通过添加草酸盐或磷酸盐负荷诱导尿液结晶。收集晶体，洗涤，并通过扫描电子显微镜、X射线衍射和能量色散X射线微分析进行分析。通过用乙二胺四乙酸（EDTA）脱矿获得基质蛋白，通过聚丙烯酰胺凝胶电泳进行分析，并通过蛋白质印迹技术进行鉴定。在CaOx和CaP晶体基质之间以及从正常人和结石形成者尿液中获得的晶体基质之间未检测到显著差异。在正常人和结石形成者尿液中诱导生成的CaOx和CaP晶体基质中均鉴定出白蛋白（AB）、α-间抑制因子（IalphaI）相关蛋白、α-1微球蛋白（α-1 m）、骨桥蛋白（OPN）、凝血酶原（PT）相关蛋白和Tamm-Horsfall蛋白（THP）。AB、PT相关蛋白和OPN是主要成分。其他蛋白质含量较少但可检测到。然而，CaP晶体基质含有大量THP。此外，CaP晶体含有的蛋白质明显多于CaOx晶体。尿液的离心和/或过滤导致尿液中许多高分子量蛋白质减少，包括THP、AB和OPN。
