Haghi Mehra, Hittinger Marius, Zeng Qingxiang, Oliver Brian, Traini Daniela, Young Paul M, Huwer Hanno, Schneider-Daum Nicole, Lehr Claus-Michael
†Drug Delivery, Helmholtz Institute for Pharmaceutical Research Saarland (HIPS), Helmholtz Centre for Infection Research (HZI), 66123 Saarbrücken, Germany.
‡School of Pharmacy, Graduate School of Health, The University of Technology Sydney, New South Wales 2007, Australia.
Mol Pharm. 2015 Aug 3;12(8):2625-32. doi: 10.1021/acs.molpharmaceut.5b00124. Epub 2015 Jul 6.
The aim of this study was to investigate the changes in transport and effectiveness of salbutamol sulfate (SAL) and budesonide (BD) following stimulation with transforming growth factor-β (TGF-β) in mono- and coculture models of bronchial and alveolar epithelium. Primary bronchial and alveolar epithelial cells, grown at air interface on filters, either as monocultures or in coculture with airway smooth muscle cells or alveolar macrophages, respectively, were stimulated with TGF-β. The biological response was modulated by depositing aerosolized SAL and BD on bronchial and alveolar models, respectively. Barrier integrity, permeability to fluorescein-Na, transport of the deposited drug, and the pharmacological response to SAL (cAMP and IL-8 levels) or BD (IL-6 and -8 levels) were measured. While stimulation with TGF-β did not have any significant effect on the transepithelial electrical resistance and permeability to fluorescein-Na in mono- and coculture models, transport of SAL and BD were affected in cultures from some of the patients (6 out of 12 for bronchial and 2 out of 4 for alveolar cells). The bronchial coculture showed a better responsiveness to SAL in terms of cAMP release than the monoculture. In contrast, the difference between alveolar mono- and cocultures to TGF-β mediated interleukin release and its modulation by BD was less pronounced. Our data point to intrinsic differences in the transport of, and responsiveness to, SAL and BD when epithelial cell cultures originate from different patients. Moreover, if the biological responses (e.g., IL-8, cAMP) involve communication between different cell types, coculture models are more relevant to measure such effects than monocultures.
本研究的目的是在支气管和肺泡上皮的单培养和共培养模型中，研究转化生长因子-β（TGF-β）刺激后硫酸沙丁胺醇（SAL）和布地奈德（BD）的转运及有效性变化。将原代支气管和肺泡上皮细胞接种于滤器上的气液界面，分别进行单培养或与气道平滑肌细胞或肺泡巨噬细胞共培养，然后用TGF-β刺激。通过分别将雾化的SAL和BD沉积在支气管和肺泡模型上来调节生物学反应。测量屏障完整性、对荧光素钠的通透性、沉积药物的转运以及对SAL（cAMP和IL-8水平）或BD（IL-6和-8水平）的药理反应。虽然在单培养和共培养模型中，TGF-β刺激对跨上皮电阻和对荧光素钠的通透性没有显著影响，但一些患者的培养物中SAL和BD的转运受到了影响（支气管细胞12例中有6例，肺泡细胞4例中有2例）。支气管共培养在cAMP释放方面对SAL的反应性比单培养更好。相比之下，肺泡单培养和共培养对TGF-β介导的白细胞介素释放及其受BD调节的差异不太明显。我们的数据表明，当上皮细胞培养物来自不同患者时，SAL和BD在转运和反应性方面存在内在差异。此外，如果生物学反应（如IL-8、cAMP）涉及不同细胞类型之间的通讯，那么共培养模型比单培养模型更适合测量此类效应。
