Albuminuria is not associated with elevated urinary vesicle concentration but can confound nanoparticle tracking analysis
Albuminuria is not associated with elevated urinary vesicle concentration but can confound nanoparticle tracking analysis
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DOI：
10.1111/nep.12867
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发表时间：
2017-11-01
期刊：
影响因子：
2.5
通讯作者：
Gleadle, Jonathan M.
中科院分区：
文献类型：
--
作者：
McNicholas, Kym;Li, Jordan Y. Z.;Gleadle, Jonathan M.
细胞外囊泡，如外泌体，存在于尿液中，有报道称其在细胞间信号传导和诊断中发挥作用。然而，在蛋白尿性肾病中尿囊的浓度和特征改变的程度还没有得到很好的表征。在这项研究中，我们研究了白蛋白尿urinase.MethodsVesicles的细胞外囊泡的数量和特点，从32例患者的尿液中分离出不同水平的白蛋白尿，使用ultracentragation和密度梯度纯化，并进行了检查，使用纳米粒子跟踪分析，免疫印迹和透射电子显微镜。在这些尿制剂中的颗粒的大小配置文件进行了比较与含白蛋白solutions.ResultsOverall，有释放到尿蛋白的囊泡的数量，或特性，没有实质性差异。含白蛋白的溶液的分析显示了外泌体样大小的颗粒，表明这种颗粒可以在标准纳米颗粒跟踪分析中模拟外泌体。蛋白尿的白蛋白和IgG耗尽导致纳米粒子跟踪analysis.ConclusionThere检测到的颗粒浓度大幅降低尿蛋白尿患者的尿囊浓度没有增加。此外，这些结果表明，需要谨慎的解释纳米粒子跟踪分析的生物液体中含有蛋白质的囊泡浓度和复杂的制备方法，从urina.Summary的囊泡纯化的数量和特点的尿囊泡的一瞥检查表明，白蛋白尿和正常尿之间没有显着差异。
AimExtracellular vesicles, such as exosomes, are present in urine with reports of roles in intercellular signalling and diagnostic utility. However, the extent to which the concentration and characteristics of urinary vesicles are altered in albuminuric renal disease has not been well characterized. In this study, we examined the number and characteristics of extracellular vesicles in albuminuric urine.MethodsVesicles were isolated from the urine of 32 patients with varying levels of albuminuria using ultracentrifugation and density gradient purification and were examined using nanoparticle tracking analysis, immunoblotting and transmission electron microscopy. The size profile of particles in these urine preparations was compared with albumin-containing solutions.ResultsOverall, there were no substantial differences in the number, or characteristics, of vesicles released into proteinuric urine. Analysis of albumin-containing solutions showed particles of exosome-like size, suggesting that such particles can mimic exosomes in standard nanoparticle tracking analysis. Albumin and IgG depletion of proteinuric urine resulted in a substantial reduction in the concentration of particles detected by nanoparticle tracking analysis.ConclusionThere was no increase in urinary vesicle concentration in patients with albuminuria. Furthermore, these results demonstrate the need for cautious interpretation of nanoparticle tracking analysis of vesicle concentration in biological fluids containing protein and for sophisticated preparative methods in vesicle purification from urine.Summary at a Glance Examination of the number and characteristics of urinary vesicles showed no significant differences between albuminuric and normal urine.