Document Detail


Characterization of protein factor(s) in rat bronchoalveolar lavage fluid that enhance insulin transport via transcytosis across primary rat alveolar epithelial cell monolayers.
MedLine Citation:
PMID:  18406118     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The aim of this study was to characterize factor(s) in rat bronchoalveolar lavage fluid (BALF) that enhance(s) insulin transport across primary rat alveolar epithelial cell monolayers (RAECM) in primary culture. BALF was concentrated 7.5-fold using the Centricon device and the retentate was used to characterize the factor(s) involved in enhancing apical-to-basolateral transport of intact 125I-insulin across various epithelial cell monolayers. These factor(s) enhanced transport of intact insulin across type II cell-like RAECM (3-fold increase) and type I cell-like RAECM (2-fold increase), but not across Caco-2 or MDCK cell monolayers. The insulin transport-enhancing factor(s) were temperature- and trypsin-sensitive. The mechanism of enhancement did not seem to involve paracellular transport or fluid-phase endocytosis, since fluxes of sodium fluorescein and FITC-dextran (70 kDa) were not affected by the factor(s) in the apical bathing fluid. BALF enhancement of intact 125I-insulin transport was abolished at 4 degrees C and in the presence of monensin, suggesting involvement of transcellular pathways. Sephacryl S-200 purification of BALF retentate, followed by LC-MS/MS, indicated that the high molecular weight (>100 kDa) fractions (which show some homology to alpha-1-inhibitor III, murinoglobulin gamma 2, and pregnancy-zone protein) appear to facilitate transcellular transport of insulin across RAECM.
Authors:
Rana Bahhady; Kwang-Jin Kim; Zea Borok; Edward D Crandall; Wei-Chiang Shen
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2008-02-06
Journal Detail:
Title:  European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft für Pharmazeutische Verfahrenstechnik e.V     Volume:  69     ISSN:  0939-6411     ISO Abbreviation:  Eur J Pharm Biopharm     Publication Date:  2008 Aug 
Date Detail:
Created Date:  2008-08-04     Completed Date:  2008-10-30     Revised Date:  2013-06-05    
Medline Journal Info:
Nlm Unique ID:  9109778     Medline TA:  Eur J Pharm Biopharm     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  808-16     Citation Subset:  IM    
Affiliation:
Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90089-9121, USA.
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MeSH Terms
Descriptor/Qualifier:
Absorption
Animals
Biological Transport, Active / drug effects
Bronchoalveolar Lavage Fluid / chemistry*
Caco-2 Cells
Cell Line
Cells, Cultured
Databases, Genetic
Dextrans
Dogs
Endocytosis / drug effects*
Epithelial Cells / drug effects,  metabolism*
Fluorescein-5-isothiocyanate / analogs & derivatives
Fluorescent Dyes
Humans
Insulin / metabolism*
Mass Spectrometry
Peptides / isolation & purification,  pharmacology*
Pulmonary Alveoli / cytology,  metabolism*
Rats
Rats, Sprague-Dawley
Stimulation, Chemical
Temperature
Trypsin / pharmacology
Grant Support
ID/Acronym/Agency:
HL38578/HL/NHLBI NIH HHS; HL38621/HL/NHLBI NIH HHS; HL38658/HL/NHLBI NIH HHS; HL62569/HL/NHLBI NIH HHS; HL64365/HL/NHLBI NIH HHS; R01 HL064365/HL/NHLBI NIH HHS
Chemical
Reg. No./Substance:
0/Fluorescent Dyes; 0/Insulin; 0/Peptides; 0/fluorescein isothiocyanate dextran; 3326-32-7/Fluorescein-5-isothiocyanate; 9004-54-0/Dextrans; EC 3.4.21.4/Trypsin
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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