Paracellular calcium flux across Caco-2 cell monolayers: Effects of individual amino acids

High-protein diets are generally considered beneficial for calcium (Ca) economy and bone health. Improved intestinal Ca absorption efficiency may be one mechanism by which higher-protein diets affect Ca homeostasis and bone health. The signaling pathways and individual amino acids (AA) responsible for this effect have not been fully elucidated and may involve the transcellular pathway, paracellular pathway or a combination. The primary aim of this study was to investigate whether a mixture of AA and/or functionally distinct individual AA directly affect paracellular Ca absorption across an intestinal epithelial cell model (Caco-2 Bbe). Using Ussing chambers, we examined the effect of six treatments - vehicle (Veh), 80 mM raffinose (Raf; positive control), 2× mixed AA(2×AA, twice the concentration in standard growth media), the branched-chain amino acid leucine (2-10 mM Leu), the aromatic amino acid phenylalanine (2-10 mM Phe) and the dibasic amino acid lysine (2-10 mM Lys) - on Ca flux. Leu (5 mM) increased Ca flux by 38% (+122 nmol Ca/cm²/h, P<.001) as compared to Veh, while 10 mM Phe reduced Ca flux. No other differences were observed. Leu increased Ca flux through cellular redistribution of the Ca permissive channel Cldn-2 to the tight junction membrane (P<.05). Inhibition of mTORC1 signaling did not abrogate the effect of Leu on Cldn-2 localization, indicating a non-mTORC1-dependent signaling pathway is involved. These data indicate that Leu may improve Ca absorption in a cell model, potentially contributing to the observed benefits of higher-protein diets on bone health in humans.