Evidence for predominantly paracellular transport of thyrotropin-releasing hormone across CACO-2 cell monolayers

Uptake, transport, and metabolism of tripeptide thyrotropin-releasing hormone were examined in the human intestinal epithelial cell line, Caco-2. A linear relationship between rate and concentration was observed for both the uptake and the transport of thyrotropin-releasing hormone across Caco-2 cell monolayers. Transport of thyrotropin-releasing hormone was not affected by the presence of dipeptide glycylsarcosine, amino acid glycine, tripeptide thyrotropin-releasing hormone free acid as well as active transport inhibitors 2,4-dinitrophenol, sodium azide, ouabain, and amiloride. There was no formation of metabolites during the course of thyrotropin-releasing hormone transport across Caco-2 cells. Incubation of Caco-2 cell homogenate with thyrotropin-releasing hormone, however, showed a time-dependent hydrolysis of thyrotropin-releasing hormone and the formation of thyrotropin-releasing hormone free acid. Increased rate of transport in the presence of EDTA indicates a paracellular passive diffusion as the major route for the transport of TRH. The hydrolytic enzyme present in Caco-2 cells appeared to have little or no access to TRH during the transcellular transport across Caco-2 cell monolayers.