PURPOSE: Examination of the contribution of functional P-glycoprotein (P-gp), an ATP-dependent efflux pump, in blood-aqueous barrier in rabbits. METHODS: Rhodamine-123 (Rho-123), a P-gp substrate, was administered intravenously via the marginal ear vein of rabbits. Rhodamine B (Rho-B), an analogue of Rho-123, was also injected with the same dose, as a reference compound. Quinidine at different concentrations was applied topically to the corneal surface by eye drops prior to the intravenous administration of a Rho compound. The aqueous distribution (a ratio of concentration in aqueous humor to that in plasma) of these Rho compounds was analyzed in relation to the aqueous concentration of quinidine. Transport study across Caco-2 cell monolayers was carried out to examine the involvement of P-gp in Rho-B transport. RESULTS: It was proved that Rho-B is not a P-gp substrate by a transport study across Caco-2 cell monolayers, in contrast to Rho-123 (a P-gp substrate). The aqueous distribution of Rho-123 given intravenously was significantly lower than that of Rho-B. Topical quinidine (a P-gp inhibitor) markedly increased the aqueous distribution of Rho-123, depending on the aqueous concentrations of quinidine, though it did not affect the aqueous distribution of Rho-B. CONCLUSIONS: The contribution of functional P-gp in blood-aqueous barrier was clearly demonstrated by analyzing the aqueous distribution of Rho-123 in the presence or absence of quinidine. These experiments only allow us to address one part of the blood-aqueous barrier, the capillary endothelium, and, to do so by using different substrates for P-gp, a sort of chemical analogy with the presumed blood-aqueous barrier across capillary endothelia. The alteration of P-gp function by pharmacotherapy or in pathological state should be considered in the ophthalmic medical treatment.
PURPOSE: Examination of the contribution of functional P-glycoprotein (P-gp), an ATP-dependent efflux pump, in blood-aqueous barrier in rabbits. METHODS:Rhodamine-123 (Rho-123), a P-gp substrate, was administered intravenously via the marginal ear vein of rabbits. Rhodamine B (Rho-B), an analogue of Rho-123, was also injected with the same dose, as a reference compound. Quinidine at different concentrations was applied topically to the corneal surface by eye drops prior to the intravenous administration of a Rho compound. The aqueous distribution (a ratio of concentration in aqueous humor to that in plasma) of these Rho compounds was analyzed in relation to the aqueous concentration of quinidine. Transport study across Caco-2 cell monolayers was carried out to examine the involvement of P-gp in Rho-B transport. RESULTS: It was proved that Rho-B is not a P-gp substrate by a transport study across Caco-2 cell monolayers, in contrast to Rho-123 (a P-gp substrate). The aqueous distribution of Rho-123 given intravenously was significantly lower than that of Rho-B. Topical quinidine (a P-gp inhibitor) markedly increased the aqueous distribution of Rho-123, depending on the aqueous concentrations of quinidine, though it did not affect the aqueous distribution of Rho-B. CONCLUSIONS: The contribution of functional P-gp in blood-aqueous barrier was clearly demonstrated by analyzing the aqueous distribution of Rho-123 in the presence or absence of quinidine. These experiments only allow us to address one part of the blood-aqueous barrier, the capillary endothelium, and, to do so by using different substrates for P-gp, a sort of chemical analogy with the presumed blood-aqueous barrier across capillary endothelia. The alteration of P-gp function by pharmacotherapy or in pathological state should be considered in the ophthalmic medical treatment.