Characterization of the Aquaporin-9 Inhibitor RG100204 In Vitro and in db/db Mice.
Characterization of the Aquaporin-9 Inhibitor RG100204 In Vitro and in db/db Mice.
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作者：
水通道蛋白9（AQP 9）是甘油等中性小分子溶质跨膜扩散的促进剂。水通道蛋白家族蛋白的特异性抑制剂的鉴定一直是困难的，这是由于13种人类同种型之间的高度序列相似性，以及由于允许抑制剂结合的有限的通道表面积。迄今为止描述的几种AQP 9抑制剂分子不适合于体内实验。我们现在描述一种新的小分子AQP 9抑制剂，RG 100204在基于细胞的钙黄绿素淬灭试验，并通过停流光散射记录AQP 9的渗透性在蛋白脂质体的表征。此外，我们研究了RG 100204对小鼠甘油代谢的影响。在基于细胞的试验中，RG 100204以相似的高效力（约5 × 10−8 M）阻断AQP 9水渗透性和甘油渗透性。在蛋白脂质体中证实了RG 100204对AQP 9通道的阻断。经口灌胃给予db/db小鼠RG 100204后，观察到血浆甘油呈剂量依赖性升高。降糖作用无统计学显著性。这些实验确立了RG 100204作为AQP 9通道的直接阻断剂，并建议将其用作AQP 9功能体内实验的实验工具。
Aquaporin-9 (AQP9) is a facilitator of glycerol and other small neutral solute transmembrane diffusion. Identification of specific inhibitors for aquaporin family proteins has been difficult, due to high sequence similarity between the 13 human isoforms, and due to the limited channel surface areas that permit inhibitor binding. The few AQP9 inhibitor molecules described to date were not suitable for in vivo experiments. We now describe the characterization of a new small molecule AQP9 inhibitor, RG100204 in cell-based calcein-quenching assays, and by stopped-flow light-scattering recordings of AQP9 permeability in proteoliposomes. Moreover, we investigated the effects of RG100204 on glycerol metabolism in mice. In cell-based assays, RG100204 blocked AQP9 water permeability and glycerol permeability with similar, high potency (~5 × 10−8 M). AQP9 channel blocking by RG100204 was confirmed in proteoliposomes. After oral gavage of db/db mice with RG100204, a dose-dependent elevation of plasma glycerol was observed. A blood glucose-lowering effect was not statistically significant. These experiments establish RG100204 as a direct blocker of the AQP9 channel, and suggest its use as an experimental tool for in vivo experiments on AQP9 function.
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影响因子：
5.6
作者：
Ampah-Korsah H;Anderberg HI;Engfors A;Kirscht A;Norden K;Kjellstrom S;Kjellbom P;Johanson U
通讯作者：
Johanson U
影响因子：
4
作者：
Calamita G;Perret J;Delporte C
通讯作者：
Delporte C
影响因子：
7.3
作者：
通讯作者：
--
影响因子：
5.5
作者：
通讯作者：
--
影响因子：
4.8
作者：
Tsukaguchi, H;Shayakul, C;Hediger, MA
通讯作者：
Hediger, MA