Kinetics of phosphorus absorption and expressions of related transporters in primary cultured duodenal epithelial cells of chick embryos.

Abstract

Two experiments were conducted to investigate the kinetics of phosphorus (P) absorption and expressions of type IIb sodium-dependentphosphate cotransporter (NaP-IIb), inorganic phosphate transporters 1 and 2 (PiT-1 and PiT-2) in primary cultured duodenal epithelial cells of chick embryos. In experiment 1, the P absorptions across duodenal epithelial cell monolayers at different incubation time points (0, 10, 20, 40, 60, 80, 100 and 120min) were compared. In experiment 2, the kinetics of P absorption was performed at 40min after incubation of duodenal epithelial cells with the media containing 0, 0.75, 1.5, 3.0, 6.0, 12.0, 24.0 and 48.0mmol P/L as KH2 PO4 , and the mRNA and protein expression levels of NaP-IIb, PiT-1 and PiT-2 in duodenal epithelial cells with the media containing 0, 6.0 and 48.0mmol P/L were determined at 87min after incubation. The results from experiment 1 showed that the P absorption increased linearly (p<.0001) from 0 to 80min and the fastest increase occurred at 40min; the asymptotic model was shown to have the best fit degree, and the optimal incubation time for saturable P absorption was determined to be 87min. The kinetic curves of P absorption from experiment 2 demonstrated that P absorption was a mixed process of a non-saturable diffusion plus a saturable carrier-mediated transport across the duodenal epithelial cells. The high P concentration (48.0mmol/L) decreased (p<.05) NaP-IIb and PiT-1 mRNA and protein levels and increased (p<.0001) PiT-2 mRNA level. These results indicated that the P absorption across primary cultured duodenal epithelial cell monolayers of chick embryos was a mixed process of a non-saturable diffusion plus a saturable carrier-mediated transport and could be restricted by reducing the NaP-IIb and PiT-1 expressions while increasing the PiT-2 expression at a high P concentration.