title: Enhancement effect of poly(amino acid)s on insulin uptake in alveolar epithelial cells.
abstract: In this study, we elucidated the effect of poly(amino acid)s such as poly-L-ornithine (PLO) on FITC-insulin uptake in cultured alveolar type II epithelial cells, RLE-6TN. FITC-insulin uptake by RLE-6TN cells as well as its cell surface binding was markedly increased by PLO without cytotoxicity. The uptake of FITC-insulin in the presence of PLO was shown to be mediated by endocytosis, but in contrast to the uptake in the absence of PLO, the contribution of macropinocytosis emerged. Colocalization of FITC-insulin and LysoTracker Red was observed by confocal laser scanning microscopy both in the absence and presence of PLO, indicating that FITC-insulin was partly targeted to lysosomes in the cells and degraded. The half-life of the intracellular degradation of FITC-insulin was, however, prolonged by the presence of PLO. PLO also stimulated the uptake of other FITC-labeled compounds. Among them, the enhancement effects of PLO on FITC-albumin and FITC-insulin uptake were prominent. The effect of PLO on insulin absorption was also examined in in-vivo pulmonary administration in rats, and co-administration of PLO enhanced the hypoglycemic action of insulin. These findings suggest that co-administration of poly(amino acid)s such as PLO is a useful strategy for enhancing insulin uptake by alveolar epithelial cells and subsequent absorption from the lung.
authors.affiliation: Department of Pharmaceutics and Therapeutics, Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Oda
authors.forename: Ryoko
authors.initials: R
authors.lastname: Yumoto
authors.forename: Junya
authors.initials: J
authors.lastname: Nagai
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Katayama
authors.forename: Mikihisa
authors.initials: M
authors.lastname: Takano
mesh_terms: D000042:Absorption; D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D004347:Drug Interactions; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000493:pharmacokinetics; D006207:Half-Life; D007004:Hypoglycemic Agents / Q000493:pharmacokinetics; D007328:Insulin / Q000031:analogs & derivatives / Q000378:metabolism / Q000493:pharmacokinetics*; D008247:Lysosomes / Q000378:metabolism; D008297:Male; D010455:Peptides / Q000494:pharmacology*; D010873:Pinocytosis / Q000502:physiology; D011650:Pulmonary Alveoli / Q000166:cytology / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000596:Amino Acids; D007004:Hypoglycemic Agents; D007328:Insulin; D010455:Peptides; C031573:insulin, fluorescein-isothiocyanated-; C008973:polyornithine; D016650:Fluorescein-5-isothiocyanate