Transport of thiamine in human intestine: mechanism and regulation in intestinal epithelial cell model Caco-2
| Autor: | Stanley A. Rubin, Hamid M. Said, Nabendu S. Chatterjee, Pradeep K. Dudeja, Alvaro Ortiz, Chandira K. Kumar |
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| Rok vydání: | 1999 |
| Předmět: |
Vitamin
Time Factors Physiology chemistry.chemical_element Calcium chemistry.chemical_compound Cations medicine Humans Thiamine Intestinal Mucosa Thiamine transport Osmolar Concentration Sodium Temperature Biological Transport Cell Biology Hydrogen-Ion Concentration Membrane transport Epithelium B vitamins medicine.anatomical_structure Biochemistry chemistry Caco-2 Caco-2 Cells Protein Kinases |
| Zdroj: | American Journal of Physiology-Cell Physiology. 277:C645-C651 |
| ISSN: | 1522-1563 0363-6143 |
| DOI: | 10.1152/ajpcell.1999.277.4.c645 |
| Popis: | The present study examined the intestinal uptake of thiamine (vitamin B1) using the human-derived intestinal epithelial cells Caco-2 as an in vitro model system. Thiamine uptake was found to be 1) temperature and energy dependent and occurred with minimal metabolic alteration; 2) pH sensitive; 3) Na+independent; 4) saturable as a function of concentration with an apparent Michaelis-Menten constant of 3.18 ± 0.56 μM and maximal velocity of 13.37 ± 0.94 pmol ⋅ mg protein−1⋅ 3 min−1; 5) inhibited by the thiamine structural analogs amprolium and oxythiamine, but not by unrelated organic cations tetraethylammonium, N-methylnicotinamide, and choline; and 6) inhibited in a competitive manner by amiloride with an inhibition constant of 0.2 mM. The role of specific protein kinase-mediated pathways in the regulation of thiamine uptake by Caco-2 cells was also examined using specific modulators of these pathways. The results showed possible involvement of a Ca2+/calmodulin (CaM)-mediated pathway in the regulation of thiamine uptake. No role for protein kinase C- and protein tyrosine kinase-mediated pathways in the regulation of thiamine uptake was evident. These results demonstrate the involvement of a carrier-mediated system for thiamine uptake by Caco-2 intestinal epithelial cells. This system is Na+independent and is different from the transport systems of organic cations. Furthermore, a CaM-mediated pathway appears to play a role in regulating thiamine uptake in these cells. |
| Databáze: | OpenAIRE |
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