School of Public Health, Sun Yat-sen University, 74 Zhongshan Road 2, Guangzhou 510080, PR China.
Toxicol Lett. 2012 Aug 3;212(3):307-14. doi: 10.1016/j.toxlet.2012.05.017. Epub 2012 Jun 9.
Melamine can cause urinary stones related to nephropathy of the kidney and hyperplasia or carcinoma of the bladder, but the mechanism of stone formation is not well understood. In this study, male rats were administered melamine for thirteen weeks to establish melamine bladder stone models and the stones were analysed by Fourier transform infrared (FTIR) spectroscopy, powder X-ray diffraction (XRD), energy dispersive X-ray (EDX) spectroscopy, sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), liquid chromatography/mass spectrometry/mass spectrometry (LC-MS/MS) and western blot, respectively, for the composition and proteome, and to explore the implication of proteins for stone formation. The results showed bladder stones were composed of predominant melamine and a few amount of proteins. The proteins had a wide range of molecular weights and 1051 proteins were identified. Gene Ontology (GO) classification of the identified proteins showed most proteins were from injured cells, involved in various metabolic processes and had binding functions. Of the identified proteins, there were a few inflammatory proteins and urinary proteins. Physicochemical characteristics of the identified proteins showed that 67.1% proteins' isoelectric points (pI) value was below 7.0, 91.1% proteins' grand average of hydropathicity (GRAVY) scores were below 0 and nearly half of the proteins were stable. Our data indicated proteins might play an important role in melamine bladder stone formation.
三聚氰胺可导致与肾病相关的肾结石以及膀胱增生或癌,但其结石形成机制尚不清楚。本研究通过给雄性大鼠十三周三聚氰胺灌胃建立三聚氰胺膀胱结石模型,采用傅里叶变换红外(FTIR)光谱、粉末 X 射线衍射(XRD)、能谱(EDX)、十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)、液相色谱/质谱/质谱联用(LC-MS/MS)和蛋白质印迹法分别对结石成分和蛋白质组进行分析,以探讨蛋白质与结石形成的关系。结果表明,膀胱结石主要由三聚氰胺和少量蛋白质组成。这些蛋白质的分子量范围很广,鉴定出 1051 种蛋白质。GO 分类表明,鉴定出的蛋白质大多数来源于受损细胞,参与多种代谢过程,具有结合功能。其中,有少量炎症蛋白和尿蛋白。鉴定出的蛋白质的理化特性表明,67.1%的蛋白质等电点(pI)值低于 7.0,91.1%的蛋白质平均疏水性(GRAVY)值低于 0,近一半的蛋白质稳定。这些数据表明蛋白质可能在三聚氰胺膀胱结石形成中发挥重要作用。