Tawada T, Fujita K, Sakakura T, Shibutani T, Nagata T, Iguchi M, Kohri K
Department of Urology, Nagoya City University School of Medicine, Nagoya, Japan.
Urol Res. 1999 Aug;27(4):238-42. doi: 10.1007/s002400050116.
We recently reported that osteopontin (OPN) and calprotectin (CPT) are present in the matrix of urinary calcium stones, and that OPN mRNA is expressed in the renal distal tubular cells. In the present study, we examined the immunohistochemical distributions of OPN and CPT in urinary stones. The stones used in this study were passed spontaneously from the upper urinary tract. One half of each of the stones was analyzed with an infrared spectrophotometer, and were shown to be comprised of calcium oxalate, calcium phosphate, uric acid and cystine. The other half of each stone was immersed in tetrasodium ethylenediamine-tetraacetate (EDTA) solution. The half-stones were embedded in paraffin and cut into 5-microm sections. The avidin-biotin-peroxidase complex technique was employed. A monoclonal antibody to human milk-derived OPN and a monoclonal antibody to human granulocyte-derived CPT were used as primary antibodies. The immunochemical study using the OPN and CPT antibodies showed positive staining of the matrix of the urinary calcium stones. The stones showed staining in two distinct zones: a core area was stained with randomly aggregated OPN and CPT, and peripheral layers were stained in concentric circles. On the basis of our observations, it is reasonable to presume that OPN and CPT play roles as the matrix in the structure of urinary calcium stones.
我们最近报道，骨桥蛋白（OPN）和钙卫蛋白（CPT）存在于尿钙结石基质中，且OPN信使核糖核酸在肾远曲小管细胞中表达。在本研究中，我们检测了OPN和CPT在尿结石中的免疫组织化学分布。本研究中使用的结石是从上尿路自然排出的。每块结石的一半用红外分光光度计进行分析，结果显示其由草酸钙、磷酸钙、尿酸和胱氨酸组成。每块结石的另一半浸入乙二胺四乙酸四钠（EDTA）溶液中。将半块结石用石蜡包埋并切成5微米厚的切片。采用抗生物素蛋白-生物素-过氧化物酶复合物技术。使用抗人乳源OPN单克隆抗体和抗人粒细胞源CPT单克隆抗体作为一抗。使用OPN和CPT抗体进行的免疫化学研究显示尿钙结石基质呈阳性染色。结石在两个不同区域呈现染色：核心区域被随机聚集的OPN和CPT染色，周边层呈同心圆染色。基于我们的观察，推测OPN和CPT在尿钙结石结构中作为基质发挥作用是合理的。
