Demonstration of ATP-dependent, transcellular transport of lipid across the lymphatic endothelium using an in vitro model of the lacteal.

Purpose: The lymphatic system plays crucial roles in tissue fluid balance, trafficking of immune cells, and the uptake of dietary lipid from the intestine. Given these roles there has been an interest in targeting lymphatics through oral lipid-based formulations or intradermal delivery of drug carrier systems. However the mechanisms regulating lipid uptake by lymphatics remain unknown. Thus we sought to modify a previously developed in vitro model to investigate the role of ATP in lipid uptake into the lymphatics.

Methods: Lymphatic endothelial cells were cultured on a transwell membrane and the effective permeability to free fatty acid and Caco-2 cell-secreted lipid was calculated in the presence or absence of the ATP inhibitor sodium azide.

Results: ATP inhibition reduced Caco-2 cell-secreted lipid transport, but not dextran transport. FFA transport was ATP-dependent primarily during early periods of ATP inhibition, while Caco-2 cell-secreted lipid transport was lowered at all time points studied. Furthermore, the transcellular component of transport was highly ATP-dependent, a mechanism not observed in fibroblasts, suggesting these mechanisms are unique to lymphatics. Total transport of Caco-2 cell-secreted lipid was dose-dependently reduced by ATP inhibition, and transcellular lipoprotein transport was completely attenuated.

Conclusion: The transport of lipid across the lymphatic endothelium as demonstrated with this in vitro model occurs in part by an ATP-dependent, transcellular route independent of passive permeability. It remains to be determined the extent that this mechanism exists in vivo and future work should be directed in this area.