Assessment of transport rates of proteins and peptides across primary human alveolar epithelial cell monolayers
Assessment of transport rates of proteins and peptides across primary human alveolar epithelial cell monolayers
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DOI：
10.1016/j.ejps.2006.02.002
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发表时间：
2006-06-01
影响因子：
4.6
通讯作者：
Ehrhardt, Carsten
中科院分区：
文献类型：
--
作者：
Bur, Michael;Huwer, Hanno;Ehrhardt, Carsten
在这项研究中，我们研究了双向通量（即，在吸收和分泌方向）的人血清蛋白[白蛋白（HSA）、转铁蛋白（TF）和免疫球蛋白G（IgG）]和具有潜在治疗相关性的肽/蛋白[胰岛素（INS）、胰高血糖素样肽-1（GLP-1）、生长激素（GH）和甲状旁腺激素（PTH）]穿过原代培养的人肺泡上皮细胞（hAEpC）的紧密单层。表观渗透系数（P-app; x 10（-7）cm/s，平均值+/- S.D.）对于GLP-1（6.13 +/- 0.87（吸收）对1.91 +/- 0.51（secretive）），HSA（2.45 +/- 1.02 vs 0.21 +/- 0.31），TF（0.88 +/- 0.15 vs 0.30 +/- 0.03）和IgG（0.36 +/- 0.22 vs 0.15 +/- 0.16）都具有强烈的方向依赖性，即，净吸收，而PTH（2.20 +/- 0.30 vs 1.80 +/- 0.77）、GH（8.33 +/- 1.24 vs 9.02 +/- 3.43）和INS（0.77 +/- 0.15 vs 0.72 +/- 0.36）显示无方向性。从供体和受体流体收集的测试分子的三氯乙酸沉淀分析显示出非常小的降解。这是第一次研究一系列肽和蛋白质在人肺泡上皮屏障体外模型中的渗透性数据。这些数据表明，供试物质通过人肺泡屏障时，不存在符合被动限制扩散的明显尺寸依赖性转运，部分证实了净吸收转胞吞作用。所获得的数据显着不同，从以前发表的报告，利用不同物种的单层。可以得出结论，应优先使用同源组织，以避免种属差异。(c)2006 Elsevier B. V.保留所有权利。
In this study, we investigated bi-directional fluxes (i.e., in absorptive and secretive directions) of human serum proteins [albumin (HSA), transferrin (TF), and immunoglobulin G (IgG)] and peptides/proteins of potential therapeutic relevance [insulin (INS), glucagon-like peptide-1 (GLP-1), growth hormone (GH), and parathyroid hormone (PTH)] across tight monolayers of human alveolar epithelial cells (hAEpC) in primary culture. Apparent permeability coefficients (P-app; x 10(-7) cm/s, mean +/- S.D.) for GLP-1 (6.13 +/- 0.87 (absorptive) versus 1.91 +/- 0.51 (secretive)), HSA (2.45 +/- 1.02 versus 0.21 +/- 0.31), TF (0.88 +/- 0.15 versus 0.30 +/- 0.03), and IgG (0.36 +/- 0.22 versus 0.15 +/- 0.16) were all strongly direction-dependent, i.e., net absorptive, while PTH (2.20 +/- 0.30 versus 1.80 +/- 0.77), GH (8.33 +/- 1.24 versus 9.02 +/- 3.43), and INS (0.77 +/- 0.15 versus 0.72 +/- 0.36) showed no directionality. Trichloroacetic acid precipitation analysis of tested molecules collected from donor and receiver fluids exhibited very little degradation. This is the first study on permeability data for a range of peptides and proteins across an in vitro model of the human alveolar epithelial barrier. These data indicate that there is no apparent size-dependent transport conforming to passive restricted diffusion for the tested substances across human alveolar barrier, in part confirming net absorptive transcytosis. The obtained data differ significantly from previously published reports utilising monolayers from different species. It can be concluded that the use of homologous tissue should be preferred to avoid species differences. (c) 2006 Elsevier B.V. All rights reserved.