Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat- 173234, Solan HP, India.
Int Braz J Urol. 2013 Jan-Feb;39(1):128-36. doi: 10.1590/S1677-5538.IBJU.2013.01.16.
Proteins constitute a major portion of the organic matrix of human calcium oxalate (CaOx) renal stones and the matrix is considered to be important in stone formation and growth. The present study evaluates the effect of these proteins on oxalate injured renal epithelial cells accompanied by a 2D map of these proteins.
Proteins were isolated from the matrix of kidney stones containing CaOx as the major constituent using EGTA as a demineralizing agent. The effect of more than 3kDa proteins from matrix of human renal (calcium oxalate) CaOx stones was investigated on oxalate induced cell injury of MDCK renal tubular epithelial cells. A 2D map of >3kDa proteins was also generated followed by protein identification using MALDI-TOF MS.
The >3kDa proteins enhanced the injury caused by oxalate on MDCK cells. Also, the 2D map of proteins having MW more than 3kDa suggested the abundance of proteins in the matrix of renal stone.
Studies indicate that the mixture of >3kDa proteins in the matrix of human renal stones acts as promoter of calcium oxalate crystal nucleation and growth as it augments the renal epithelial cell injury induced by oxalate. The effect of promoters masks the inhibitors in the protein mixture thereby leading to enhanced renal cell injury. 2D map throws light on the nature of proteins present in the kidney stones.
蛋白质构成了人草酸钙（CaOx）肾结石有机基质的主要部分，而基质被认为在结石形成和生长中很重要。本研究评估了这些蛋白质对草酸损伤的肾上皮细胞的影响，并绘制了这些蛋白质的 2D 图谱。
使用 EGTA 作为脱钙剂从主要成分为 CaOx 的肾结石基质中分离蛋白质。研究了来自人肾（草酸钙）CaOx 结石基质的超过 3kDa 蛋白质对 MDCK 肾小管上皮细胞草酸诱导细胞损伤的影响。还生成了 >3kDa 蛋白质的 2D 图谱，并使用 MALDI-TOF MS 进行蛋白质鉴定。
3kDa 蛋白质增强了草酸对 MDCK 细胞造成的损伤。此外，MW 大于 3kDa 的蛋白质的 2D 图谱表明肾结石基质中存在丰富的蛋白质。
研究表明，人肾结石基质中 >3kDa 蛋白质混合物作为草酸钙晶体成核和生长的促进剂，因为它增强了草酸引起的肾上皮细胞损伤。促进剂的作用掩盖了蛋白质混合物中的抑制剂，从而导致肾细胞损伤增加。2D 图谱揭示了肾结石中存在的蛋白质的性质。
