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首页> 外文期刊>American Journal of Physiology >Effect of urine fractionation on attachment of calcium oxalate crystals to renal epithelial cells: implications for studying renal calculogenesis.
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Effect of urine fractionation on attachment of calcium oxalate crystals to renal epithelial cells: implications for studying renal calculogenesis.

机译：尿液分级对草酸钙晶体附着于肾上皮细胞的影响：对研究肾结石的意义。

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Our aim was to determine whether fractionation of human urine affects the attachment of calcium oxalate monohydrate (COM) crystals to renal cells. Urine collected from six healthy subjects was fractionated into sieved (S), centrifuged (C), centrifuged and filtered (CF), or ultrafiltered (UF). Attachment of [(14)C]COM crystals to Madin-Darby canine kidney (MDCK) cells was studied after precoating the crystals or the cells with the urine fractions and by using the same fractions as the binding medium. Protein content of the fractions and precoated crystals was analyzed with SDS-PAGE and Western blotting. All urine fractions inhibited crystal attachment. When fractions from the six urine samples were used to precoat the cells, the median inhibitions of crystal adhesion ( approximately 40%) were not significantly different. Median inhibition after preincubation of crystals was the same for the S, C, and CF fractions ( approximately 40%) but significantly greater than for the UF fraction ( approximately 28%). When fractions were used as the binding medium, median inhibitions decreased from 64% in the S fraction to 47 (C), 42 (CF), and to 29% (UF). SDS-PAGE analysis showed that centrifugation and filtration reduced the amount of Tamm-Horsfall glycoprotein (THG), which was confirmed by Western blotting. Human serum albumin, urinary prothrombin fragment 1, and osteopontin, but not THG, were present in demineralized extracts of the precoated crystals. Fractionation of human urine affects the attachment of COM crystals to MDCK cells. Hence future studies investigating regulation of crystal-cell interactions should be carried out in untreated urine as the binding medium.
机译：我们的目的是确定人类尿液的分馏是否会影响草酸钙一水合物（COM）晶体与肾细胞的附着。从六个健康受试者收集的尿液分为筛分（S），离心（C），离心和过滤（CF）或超滤（UF）。 [（14）C] COM晶体与Madin-Darby犬肾（MDCK）细胞的附着在用尿液组分预涂晶体或细胞后并使用与结合介质相同的组分进行了研究。通过SDS-PAGE和蛋白质印迹分析级分和预包被的晶体的蛋白质含量。所有尿液成分均抑制晶体附着。当使用六个尿液样本中的馏分对细胞进行预涂时，晶体粘附的中位抑制作用（约40％）没有显着差异。晶体预温育后的中值抑制作用对于S，C和CF级分相同（约40％），但明显大于对UF级分（约28％）。当使用级分作为结合介质时，中位抑制率从S级分的64％降至47（C），42（CF）和29％（UF）。 SDS-PAGE分析表明，离心和过滤减少了Tamm-Horsfall糖蛋白（THG）的量，这已通过Western印迹证实。人血清白蛋白，尿液凝血酶原片段1和骨桥蛋白（而非THG）存在于预涂层晶体的脱矿物质提取物中。人尿的分级分离会影响COM晶体与MDCK细胞的附着。因此，应在未处理的尿液中作为结合介质进行有关晶体-细胞相互作用调节的进一步研究。

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