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Nutrient Interactions and Toxicity

Various Nondigestible Saccharides Open a Paracellular Calcium Transport Pathway with the Induction of Intracellular Calcium Signaling in Human Intestinal Caco-2 Cells

Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University, Sapporo, Japan

¹To whom correspondence should be addressed. E-mail: hara{at}chem.agr.hokudai.ac.jp.

Ingestion of soluble nondigestible saccharides increases calcium absorption, and it is suggested that paracellular calcium transport contributes to this effect. However, cellular mechanisms and the contribution of active transport have not been clarified. This study examined the effects of 4 nondigestible saccharides, difructose anhydride (DFA) III, DFAIV, fructooligosaccharides, and raffinose, on active and passive calcium transport, permeability of paracellular pathways, and intracellular calcium signaling in a human intestinal Caco-2 cell monolayer. Net, active, and passive calcium transport were evaluated using ⁴⁵Ca. Transepithelial electrical resistance (TEER) and transport of lucifer yellow were measured as indicators of paracellular passage in differentiated Caco-2 cell monolayers incubated with 0–100 mmol/L of the various saccharides. The changes in intracellular calcium ion concentrations ([Ca²⁺]_i) were measured by fura-2 loading before and after the addition of each saccharide (50 or 100 mmol/L). The addition of 100 mmol/L of each saccharide to the apical medium of the Caco-2 cells enhanced net calcium transport without any changes in active calcium transport. Relative TEER was dose dependently and reversibly decreased by the addition of saccharides, and the decreases in TEER were highly correlated with net calcium transport (P < 0.001). Basolateral application of the saccharides had a slight or no effect on indicators of the paracellular pathway. Each saccharide caused an immediate and dose-dependent rise in [Ca²⁺]_i in the cells. The 4 nondigestible saccharides increased net calcium transport in the cells via the paracellular route through tight junctions. The rise in [Ca²⁺]_i induced by these saccharides may be involved in the opening of tight junctions.

KEY WORDS: • nondigestible saccharide • calcium transport • intracellular calcium ion • tight junction • Caco-2 cell

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