Effect of bioflavonoids on vincristine transport across blood–brain barrier

Abstract

Several grapefruit juice bioflavonoids, including quercetin, are reported to stimulate P-glycoprotein-mediated drug efflux from cultured tumor cells. To see whether these bioflavonoids alter the permeation of vincristine across the blood–brain barrier, we conducted experiments with cultured mouse brain capillary endothelial cells (MBEC4 cells) in vitro and ddY mice in vivo. The steady-state uptake of [ 3H]vincristine by MBEC4 cells was decreased by 10 μM quercetin, but increased by 50 μM quercetin. Similarly, the in vivo brain-to-plasma concentration ratio of [ 3H]vincristine in ddY mice was decreased by coadministration of 0.1 mg/kg quercetin, but increased by 1.0 mg/kg quercetin. Kaempferol had a similar biphasic effect on the in vitro uptake of [ 3H]vincristine. Other aglycones tested (chrysin, flavon, hesperetin, naringenin) increased [ 3H]vincristine uptake in the 10–50 μM range, and glycosides (hesperidin, naringin, rutin) were without effect. We then addressed the mechanism of the concentration-dependent biphasic action of quercetin. Verapamil, a P-glycoprotein inhibitor, inhibited the efflux of [ 3H]vincristine from MBEC4 cells, while 10 μM quercetin significantly stimulated it. The uptake of [ 3H]vincristine by MBEC4 cells was increased by inhibitors of protein kinase C, but decreased by phorbol 12-myristate-13-acetate (PMA), as well as by 10 μM quercetin. The phosphorylation level of P-glycoprotein was increased in the presence of 5 μM quercetin or 100 nM PMA, but decreased by the protein kinase C inhibitor H7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine, 30 μM). We conclude that low concentrations of quercetin indirectly activate the transport of [ 3H]vincristine by enhancing the phosphorylation (and hence activity) of P-glycoprotein, whereas high concentrations of quercetin inhibit P-glycoprotein. Our results indicate that patients taking drugs which are P-glycoprotein substrates may need to restrict their intake of bioflavonoid-containing foods and beverages, such as grapefruit juice.