Chrysosplenetin inhibits artemisinin efflux in P-gp-over-expressing Caco-2 cells and reverses P-gp/MDR1 mRNA up-regulated expression induced by artemisinin in mouse small intestine.
Pharm Biol
; 55(1): 374-380, 2017 Dec.
Article
en En
| MEDLINE
| ID: mdl-27931149
ABSTRACT
CONTEXT CYP3A4 and P-gp together form a highly efficient barrier for orally absorbed drugs and always share the same substrates. Our previous work revealed that chrysosplenetin (CHR) significantly augmented the rat plasma level and anti-malarial efficacy of artemisinin (ART), partially due to the uncompetitive inhibition effect of CHR on rat CYP3A. But the impact of CHR on P-gp is still unknown. OBJECTIVE:
The present study investigates whether CHR interferes with P-gp-mediated efflux of ART and elucidates the underlying mechanism. MATERIALS ANDMETHODS:
P-gp-over-expressing Caco-2 cells were treated with ART (10 µM) or ART-CHR (12, 1020 µM) in ART bidirectional transport experiment. ART concentration was determined by UHPLC-MS/MS method. Healthy male ICR mice were randomly divided into nine groups (n = 6) including negative control (0.5% CMC-Na solution, 13 mL/kg), ART alone (40 mg/kg), verapamil (positive control, 40 mg/kg), ART-verapamil (11, 4040 mg/kg), CHR alone (80 mg/kg), ART-CHR (10.1, 404 mg/kg), ART-CHR (11, 4040 mg/kg), ART-CHR (12, 4080 mg/kg) and ART-CHR (14, 40160 mg/kg). The drugs were administrated intragastrically for seven consecutive days. MDR1 and P-gp expression levels in mice small intestine were examined by performing RT-PCR and western blot analysis. ABC coupling ATPase activity was also determined.RESULTS:
After combined with CHR (12), Papp (AP-BL) and Papp (BL-AP) of ART changed to 4.29 × 10 - 8 (increased 1.79-fold) and 2.85 × 10 - 8 cm/s (decreased 1.24-fold) from 2.40 × 10 - 8 and 3.54 × 10 - 8 cm/s, respectively. Efflux ratio (PBA/PAB) declined 2.21-fold (p < 0.01) versus to ART alone. ART significantly up-regulated both MDR1 mRNA and P-gp levels compared with vehicle, while CHR in combination ratio of 01, 0.11, 11, 21 and 41 with ART, reversed them to normal levels as well as negative control (p < 0.05). The ATPase activities in ART-CHR 14 and CHR alone groups achieved a slight increase (p < 0.05) when compared with ART alone. DISCUSSION ANDCONCLUSION:
These results confirm that CHR inhibited P-gp activity and reverse the up-regulated P-gp and MDR1 levels induced by ART. It suggested that CHR potentially can be used as a P-gp reversal agent to obstruct ART multidrug resistance.Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Flavonoides
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ARN Mensajero
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Colon
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Subfamilia B de Transportador de Casetes de Unión a ATP
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Artemisininas
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Intestino Delgado
Límite:
Animals
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Humans
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Male
Idioma:
En
Revista:
Pharm Biol
Año:
2017
Tipo del documento:
Article