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益生菌大肠杆菌菌株Nissle 1917脂多糖对5-氨基水杨酸跨Caco-2细胞单层经上皮转运的影响。

The effect of probiotic Escherichia coli strain Nissle 1917 lipopolysaccharide on the 5-aminosalicylic acid transepithelial transport across Caco-2 cell monolayers.

作者信息

Stětinová Věra, Smetanová Libuše, Kholová Dagmar, Květina Jaroslav, Svoboda Zbyněk, Zídek Zdeněk, Tlaskalová-Hogenová Helena

机构信息

Institute of Experimental Biopharmaceutics, Joint Research Centre of PRO.MED.CS Praha a.s. and the Academy of Sciences of the Czech Republic, Hradec Králové, Czech Republic.

出版信息

Gen Physiol Biophys. 2013 Sep;32(3):371-80. doi: 10.4149/gpb_2013035. Epub 2013 Jul 12.

Abstract

The object of this study was to investigate the effect of probiotic Escherichia coli strain Nissle 1917 (EcN) (i) EcN lipopolysaccharide (EcN LPS) and (ii) bacteria-free supernatant of EcN suspension (EcN supernatant) on in vitro transepithelial transport of mesalazine (5-aminosalicylic acid, 5-ASA), the most commonly prescribed anti-inflammatory drug in inflammatory bowel disease (IBD). Effect of co-administered EcN LPS (100 µg/ml) or EcN supernatant (50 µg/ml) on the 5-ASA transport (300 µmol/l) was studied using the Caco-2 monolayer (a human colon carcinoma cell line) as a model of human intestinal absorption. Permeability characteristics for absorptive and secretory transport of parent drug and its intracellularly-formed metabolite were determined. The quantification of 5-ASA and its main metabolite N-acetyl-5-amino-salicylic acid (N-Ac-5-ASA) was performed by high performance liquid chromatography. Obtained results suggest that neither EcN LPS nor EcN supernatant had effect on the total 5-ASA transport (secretory flux greater than absorptive flux) and on the transport of intracellularly formed N-Ac-5-ASA (preferentially transported in the secretory direction). The percent cumulative transport of the total 5-ASA alone or in combination with EcN LPS or EcN supernatant did not exceed 1%.

摘要

本研究的目的是调查益生菌大肠杆菌菌株Nissle 1917(EcN)(i)EcN脂多糖(EcN LPS)和(ii)EcN悬浮液的无菌上清液(EcN上清液)对美沙拉嗪(5-氨基水杨酸,5-ASA)体外跨上皮转运的影响,美沙拉嗪是炎症性肠病(IBD)中最常用的抗炎药物。使用Caco-2单层细胞(一种人结肠癌细胞系)作为人体肠道吸收模型,研究了共同给予EcN LPS(100 µg/ml)或EcN上清液(50 µg/ml)对5-ASA转运(300 µmol/l)的影响。测定了母体药物及其细胞内形成的代谢物的吸收性和分泌性转运的通透性特征。通过高效液相色谱法对5-ASA及其主要代谢物N-乙酰-5-氨基水杨酸(N-Ac-5-ASA)进行定量。获得的结果表明,EcN LPS和EcN上清液均未对总5-ASA转运(分泌通量大于吸收通量)以及细胞内形成的N-Ac-5-ASA的转运(优先沿分泌方向转运)产生影响。单独或与EcN LPS或EcN上清液联合使用时,总5-ASA的累积转运百分比均未超过1%。

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