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Am J Physiol Cell Physiol 277: C645-C651, 1999;
0363-6143/99 $5.00
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Vol. 277, Issue 4, C645-C651, October 1999

Transport of thiamine in human intestine: mechanism and regulation in intestinal epithelial cell model Caco-2

Hamid M. Said, Alvaro Ortiz, Chandira K. Kumar, Nabendu Chatterjee, Pradeep K. Dudeja, and Stanley Rubin

Veterans Affairs Medical Center, Long Beach 90822; University of California Irvine, Irvine 92717; Wadsworth Veterans Affairs Medical Center and University of California Los Angeles, Los Angeles, California 90073; and Westside Veterans Affairs Medical Center and University of Illinois-Chicago, Chicago, Illinois 60612

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.

thiamine transport; human intestinal cells; transport mechanism; transport regulation


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