Yokomizoi Kayo, Nakayama Aki, Hokazono Eisaku, Ninomiya Akiko, Miyake Ruriko, Hiratsuka Nobuo, Okuyama Mitsuhiko, Kato Yuji, Kobayashi Shizuko, Ito Yoshihisa, Shiba Kiyoko
Analytical Laboratory Chemistry, Graduate School of Allied Health Sciences, Tokyo Medical and Dental University, Bunkyo-ku, Tokyo 113-8519.
Rinsho Byori. 2005 Dec;53(12):1109-15.
In order to investigate the mechanism of urinary tract stone formation, we analyzed protein components in urine and the stone. Urinary proteins of healthy subjects and urolithic patients as well as protein components urinary tract stone of the urolithic patients were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretic patterns of urinary proteins of the patients differed from those of healthy subjects after separating protein patterns into those larger than 66kDa or smaller than 30kDa. Protein constituents of urinary tract stone were mainly separated into 18 bands ranging from 26.8 to 143 kDa. Major bands among these 18 bands differed among stones from different patients. On western blotting, the developed intensities of Tamm-Horsfall protein (THP) were fainter than those of healthy subjects. Whereas intensities of albumin (ALB) were stronger than those of healthy subjects. Moreover, blotting patterns of THP of the patients on non-reducing SDS-PAGE were obviously broad. Thus, we suggest that analysis of fractionated urinary proteins or protein components of urinary tract stone may provide a tool for monitoring the prognosis or relapse in the patients.
为了研究尿路结石形成的机制，我们分析了尿液和结石中的蛋白质成分。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）分析了健康受试者和尿路结石患者的尿蛋白以及尿路结石患者的尿路结石蛋白质成分。将患者的尿蛋白图谱分为大于66kDa或小于30kDa的图谱后，患者的尿蛋白电泳图谱与健康受试者的不同。尿路结石的蛋白质成分主要分离为18条带，范围从26.8至143kDa。这18条带中的主要条带在不同患者的结石中有所不同。在蛋白质印迹法中，Tamm-Horsfall蛋白（THP）的显色强度比健康受试者的弱。而白蛋白（ALB）的强度比健康受试者的强。此外，患者在非还原SDS-PAGE上的THP印迹图谱明显变宽。因此，我们认为分析分级尿蛋白或尿路结石的蛋白质成分可能为监测患者的预后或复发提供一种工具。
