Mamlouk Mariam, Young Paul M, Bebawy Mary, Haghi Mehra, Mamlouk Shery, Mulay Vishwaroop, Traini Daniela
Faculty of Pharmacy, University of Sydney, Sydney, New South Wales, Australia.
J Asthma. 2013 May;50(4):334-41. doi: 10.3109/02770903.2013.773518. Epub 2013 Mar 18.
The aim of this study was to characterize the permeability kinetics of salbutamol sulfate, a commonly used β2-agonist in the treatment of asthma exacerbation, across Calu-3 respiratory epithelial cell monolayers in the presence of non-steroidal anti-inflammatory drugs (NSAIDs), as they have been implicated to be able to modulate organic cation transporters (OCTs).
Calu-3 cell monolayers were grown in a liquid covered culture (LCC) configuration on 0.33 cm(2) Transwell polyester cell culture supports. Monolayers, cultured between 11 and 14 days were evaluated for epithelial resistance, tight junction integrity, and expression of OCT using Western blot analysis. The transport of salbutamol across the monolayer was studied as a function of concentration. Directional transport was investigated by assessing apical-basal (a-b) and basal-apical (b-a) directions. The influence of a non-specific OCT inhibitor (tetraethylammonium, TEA) and three NSAIDs (aspirin, ibuprofen, and indomethacin) on the uptake of salbutamol was studied.
The flux of salbutamol sulfate increased with increasing concentration before reaching a plateau, suggesting the involvement of a transport-mediated uptake mechanism. Western blot analysis detected the presence of OCT1-3 and N1 and N2 sub-types, suggesting the presence of functioning transporters. The apparent permeability (P(app)) of 0.1 mM salbutamol across the epithelial monolayer displayed directional transport in the a-b direction which was inhibited by ˜70% in the presence of TEA, suggesting OCT-mediated uptake. Likewise, the uptake of 0.1 mM salbutamol was decreased in the presence of all the three NSAIDs, supporting a mechanism whereby NSAIDs inhibit absorption of salbutamol across the bronchial epithelium via effects on the OCT transporters.
This study demonstrates that NSAIDs influence the uptake kinetics of salbutamol in an in vitro Calu-3 cell system.
本研究旨在表征硫酸沙丁胺醇（一种常用于治疗哮喘急性加重的β2激动剂）在非甾体抗炎药（NSAIDs）存在的情况下，通过Calu-3呼吸道上皮细胞单层的渗透动力学，因为它们被认为能够调节有机阳离子转运体（OCTs）。
Calu-3细胞单层在0.33平方厘米的Transwell聚酯细胞培养支持物上以液体覆盖培养（LCC）配置生长。使用蛋白质印迹分析评估培养11至14天的单层细胞的上皮电阻、紧密连接完整性和OCT的表达。研究了沙丁胺醇在单层上的转运与其浓度的关系。通过评估顶-基（a-b）和基-顶（b-a）方向来研究定向转运。研究了非特异性OCT抑制剂（四乙铵，TEA）和三种NSAIDs（阿司匹林、布洛芬和吲哚美辛）对沙丁胺醇摄取的影响。
硫酸沙丁胺醇的通量在达到平台期之前随浓度增加而增加，表明存在转运介导的摄取机制。蛋白质印迹分析检测到OCT1-3以及N1和N2亚型的存在，表明存在功能性转运体。0.1 mM沙丁胺醇在上皮单层中的表观渗透率（P(app)）在a-b方向显示出定向转运，在TEA存在下被抑制约70%，表明是OCT介导的摄取。同样，在所有三种NSAIDs存在的情况下，0.1 mM沙丁胺醇的摄取减少，支持了一种机制，即NSAIDs通过对OCT转运体的作用抑制沙丁胺醇在支气管上皮的吸收。
本研究表明，NSAIDs在体外Calu-3细胞系统中影响沙丁胺醇的摄取动力学。
