EP 80317, a CD36 selective ligand, promotes reverse cholesterol transport in apolipoprotein E-deficient mice.

AIMS: The CD36 selective ligand, EP 80317, features potent anti-atherosclerotic and hypocholesterolemic effects that are associated with an increase in macrophage cholesterol efflux through the activation of the peroxisome proliferator-activated receptor γ-liver X receptor α (LXRα)-ATP-binding cassette (ABC) transporter pathway. Cholesterol efflux is the first step of reverse cholesterol transport (RCT). However, whether EP 80317 exerts its hypocholesterolemic and anti-atherosclerotic activity through RCT in vivo has yet to be determined. In the present study, we investigated the effects of EP 80317 on RCT, in particular on macrophage-to-feces RCT and the expression of selected genes associated with hepatic cholesterol metabolism and intestinal cholesterol transport.

METHODS AND RESULTS: Reverse cholesterol transport was assessed following the intraperitoneal injection of [(3)H]-cholesterol-labelled J774 macrophages to hypercholesterolemic apoE- and apoE/CD36 double-deficient mice that had been treated for 12 weeks with EP 80317. Forty-eight hours after the administration of [(3)H]-cholesterol-labelled cells, blood, liver, intestines and feces were harvested. The radioactivity recovered in the feces (cholesterol and bile acid combined) was significantly increased by 311% (P = 0.0259) in EP 80317-treated mice compared with that found in vehicle-treated mice despite no significant change in [(3)H]-tracer recovery in plasma between groups. Whereas the mRNA levels of LXRα in the gut were significantly upregulated, mRNA and protein levels of the Niemann-Pick C1-like 1 protein (NPC1L1) transporter, a LXRα target which regulates intestinal cholesterol absorption, were downregulated in EP 80317-treated mice. In contrast, neither mRNA nor protein levels of investigated transporters and receptors were modulated in the small intestine of double-deficient mice, nor was the fecal recovery of radioactivity. No change was observed in targeted genes in liver of either apoE- or apoE/CD36 double-deficient mice after a chronic treatment with EP 80317.

CONCLUSION: This study shows that EP 80317 elicits macrophage-to-feces reverse cholesterol transport in a manner dependent on CD36 expression. This effect is associated with the upregulation of LXRα and the downregulation of NPC1L1 expression.