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MDCK 细胞分泌的基底层具有大小和电荷选择性。

Basal lamina secreted by MDCK cells has size- and charge-selective properties.

机构信息

Department of Biomedical Engineering, ND20, Cleveland Clinic, 9500 Euclid Ave., Cleveland, OH 44195, USA.

出版信息

Am J Physiol Renal Physiol. 2011 Jan;300(1):F86-90. doi: 10.1152/ajprenal.00484.2010. Epub 2010 Oct 27.

Abstract

The role electrical charge plays in determining glomerular permeability to macromolecules remains unclear. If the glomerular basement membrane (GBM) has any significant role in permselectivity, physical principles would suggest a negatively charged GBM would reject similarly charged more than neutral species. However, recent in vivo studies with negative and neutral glomerular probes showed the opposite. Whether this observation is due to unique characteristics of the probes used or is a general physiological phenomenon remains to be seen. The goal of this study was to use the basement membrane deposited by Madin-Darby canine kidney epithelial cells as a simple model of a biologically derived, negatively charged filter to evaluate size- and charge-based sieving properties. Fluorescein isothiocyanate-labeled carboxymethylated Ficoll 400 (FITC-CM Ficoll 400) and amino-4-methyl-coumarin-labeled Ficoll 400 (AMC Ficoll 400) were used as negatively charged and neutral tracer molecules, respectively, during pressure-driven filtration. Streaming potential measurement indicated the presence of fixed, negative charge in the basal lamina. The sieving coefficient for neutral Ficoll 400 decreased by ∼0.0013 for each 1-Å increment in solute radius, compared with a decrease of 0.0023 per Å for the anionic Ficoll 400. In this system, molecular charge played a significant role in determining the sieving characteristics of the membrane, pointing to solute charge as a potential contributor to GBM permselectivity.

摘要

电荷在决定大分子对肾小球的通透性方面的作用仍不清楚。如果肾小球基底膜 (GBM) 在选择性渗透方面有任何重要作用,物理原理表明带负电荷的 GBM 会排斥带相同电荷的物质多于中性物质。然而,最近用带负电和中性肾小球探针进行的体内研究结果则相反。这种观察结果是由于所使用探针的独特特性所致,还是普遍的生理现象,还有待观察。本研究的目的是使用 Madin-Darby 犬肾上皮细胞沉积的基底膜作为生物衍生的带负电荷的滤器的简单模型,评估基于大小和电荷的筛分特性。荧光素异硫氰酸酯标记的羧甲基化 Ficoll 400(FITC-CM Ficoll 400)和氨基-4-甲基香豆素标记的 Ficoll 400(AMC Ficoll 400)分别用作带负电荷和中性示踪分子,在压力驱动过滤过程中使用。流动电势测量表明基底膜中存在固定的负电荷。与带负电荷的 Ficoll 400 的筛分系数每增加 1Å 减少约 0.0013 相比,中性 Ficoll 400 的筛分系数每增加 1Å 减少约 0.0023。在这个系统中,分子电荷在决定膜的筛分特性方面起着重要作用,这表明溶质电荷可能是 GBM 选择性渗透的一个潜在贡献因素。

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