Oligopeptide Transport in Rat Lung Alveolar Epithelial Cells is Mediated by Pept2

• Published in: Pharmaceutical research 2017-12, Vol.34 (12), p.2488-2497
• Main Authors: Gukasyan, Hovhannes J., Uchiyama, Tomomi, Kim, Kwang-Jin, Ehrhardt, Carsten, Wu, Sharon K., Borok, Zea, Crandall, Edward D., Lee, Vincent H.L.
• Format: Article
• Language: English
• Subjects: Alveolar Epithelial Cells - cytology; Alveolar Epithelial Cells - metabolism; Alveoli; Animals; Biochemistry; Biological Transport; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Cells, Cultured; Drug delivery; Drug delivery systems; Drugs; Epithelial cells; Epithelium; Gene Expression; Glycine; Keratinocyte growth factor; Lungs; Male; Medical Law; Oligopeptides - metabolism; Peptide transporter; Peptides; Peptidomimetics; Pharmacology/Toxicology; Pharmacy; Polymerase chain reaction; Rats; Rats, Sprague-Dawley; Research Paper; RNA; Rodents; Sarcosine; Symporters - analysis; Symporters - genetics; Symporters - metabolism; Vehicles; Western blotting
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-017-2234-z

ABSTRACT Purpose Studies were conducted in primary cultured rat alveolar epithelial cell monolayers to characterize peptide transporter expression and function. Methods Freshly isolated rat lung alveolar epithelial cells were purified and cultured on permeable support with and without keratinocyte growth factor (KGF). Messenger RNA and protein expression of Pept1 and Pept2 in alveolar epithelial type I- and type II-like cell monolayers (±KGF, resp.) were examined by RT-PCR and Western blotting. 3 H–Glycyl-sarcosine ( 3 H–gly-sar) transmonolayer flux and intracellular accumulation were evaluated in both cell types. Results RT-PCR showed expression of Pept2, but not Pept1, mRNA in both cell types. Western blot analysis revealed presence of Pept2 protein in type II-like cells, and less in type I-like cells. Bi-directional transmonolayer 3 H–gly-sar flux lacked asymmetry in transport in both types of cells. Uptake of 3 H–gly-sar from apical fluid of type II-like cells was 7-fold greater than that from basolateral fluid, while no significant differences were observed from apical vs. basolateral fluid of type I-like cells. Conclusions This study confirms the absence of Pept1 from rat lung alveolar epithelium in vitro . Functional Pept2 expression in type II-like cell monolayers suggests its involvement in oligopeptide lung disposition, and offers rationale for therapeutic development of di/tripeptides, peptidomimetics employing pulmonary drug delivery.