Transporters involved in apical and basolateral uptake of ceftibuten into Caco-2 cells

Ceftibuten uptake from the apical and basolateral side of Caco-2 cells grown on transwells was studied. Uptake into the cells showed concentration dependent saturation. The apical transporter(s) showed a higher capacity and lower affinity for ceftibuten than the basolateral transporter(s). Uptake was inhibited in the presence of higher pH and in the presence of 2,4-dinitro phenol (DNP). A proton gradient had a greater effect on the apical than on the basolateral transporter. Glycyl proline, a dipeptide transport system (PEPT1) substrate, inhibited ceftibuten uptake into Caco-2 cells. Benzoic acid, a monocarboxylic acid (MCT) transporter substrate also exhibited a strong inhibition of ceftibuten uptake, but acetic acid had no effect. Adipic acid inhibited apical uptake of ceftibuten but had no effect on the basolateral uptake. None of the inhibitors had a significant effect on ceftibuten uptake in absence of a pH gradient. Addition of inhibitors in presence of DNP led to a greater decrease in ceftibuten uptake, when compared to the effect of DNP alone, indicating a facilitated diffusion process. These results indicate that ceftibuten uptake in Caco-2 cells involve multiple transport pathways. Apical uptake is mediated by an energy dependent carrier-mediated process and an energy independent facilitated diffusion process. The apical transport system is different from the basolateral transporter.