| Objective: To investigate the impact and its mechanism of green tea extract on rosuvastatin by pharmacokinetics studies in rats,expecting to provide useful information for rational combinative application of green tea and rosuvastatin in clinic.Method:(1)The impact of green tea extract on the pharmacokinetics of rosuvastatin in rats: Male SD rats were divided into five groups: control group of rosuvastatin(1 mg·kg-1), control group of green tea extract(200 mg·kg-1), co-administration groups of rosuvastatin(1 mg·kg-1) and green tea extract with three doses(100 mg·kg-1, 200 mg·kg-1, 500 mg·kg-1). Blood samples were withdrawn before and after dosing at different time points of 6 rats per group. Concentrations of rosuvastatin and EGCG in plasma were determined via UPLC-MS/MS method; while pharmacokinetic parameters were estimated by non-compartmental analysis using Win Nonlin software and analyzed statistically by Graph Pad software.(2)The impact of green tea extract and catechins on the uptake and transport of rosuvastatin in Caco-2 models: Caco-2 cell monolayer model was first developed and then confirmed by observing cell morphology using an inverted microscope, measuring TEER values, monitoring differentiation of alkaline phosphatase on both model sides and measuring the permeability of fluorescent yellow. UPLC-MS/MS was applied to determine the concentrations of rosuvastatin in cells and in HBSS solution. The validated Caco-2 models were applied to investigate the impacts of green tea extract and catechins on the uptake and transport of rosuvastatin.(3)The impact of green tea extract and catechins on the uptake of rosuvastatin in OATP1B1-HEK293 T cells: OATPIBl-EGFP fusion gene expression vecto was constructed using GV144 eukaryotic expression vector system carrying enhanced green fluorescent protein(EGFP) as gene transduction media. The model was validated through observing the fluorescence, detecting protein expression of EGFP by Western blot and measuring the uptake of positive substrate. UPLC-MS/MS method was applied to determine the concentrations of rosuvastatin in cells. The validated model was applied to investigate the effect of green tea extracts and catechins on the transport of rosuvastatin mediated by OATP1B1.(4)The impact of epigallocatechin gallate(EGCG) on the pharmacokinetics of rosuvastatin in rats: EGCG was administered in an amount of 4.8 mg·kg-1 in rats, which was equivalent to 200 mg·kg-1 of green tea extract, i.e. 2.4% of green tea extract. Six rats were gavaged co-administration of EGCG(4.8 mg·kg-1)with rosuvastatin(1 mg·kg-1). Concentrations of rosuvastatin and EGCG in rat plasma were determined via UPLC-MS/MS method; and pharmacokinetic parameters were estimated by non-compartmental analysis using Win Nonlin software and analyzed statistically using Graph Pad software.Results:(1)The impact of green tea extract on the pharmacokinetics of rosuvastatin in rats: After co-administrated with three different doses(100 mg·kg-1, 200 mg·kg-1, 500 mg·kg-1) green tea extract, the values of AUC0-∞ of rosuvastatin were found increased by 58.45% ± 53.04%, 85.77% ± 48.12% and 129.69% ± 69.48%, respectively; while the values of its Cmax increased by 73.96%±57.32%, 192.23%±123.56% and 152.28%±220.17%, respectively. However, there was no significant difference indicated regarding clearance values. As a result, the pharmacokinetic parameters of EGCG didn’t change significantly(p>0.05) between the co-adminstrated group of green tea extract with rosuvastatin and solo-application of green tea extract.(2)The impact of green tea extract and catechins on the uptake and transport of rosuvastatin in Caco-2 models: The green tea extract increased the uptake(0.6289±0.0748 vs 0.3245±0.0542 nmol·mg-1protein)and transport(4.3651±1.2410 x10-7 vs 2.0679±0.4370 x10-7 cm·s-1) of rosuvastatin in rats, EGCG decreased the uptake(0.2511±0.0328 vs 0.3245±0.0542 nmol·mg-1protein)and transport(1.1540±0.0762x10-7 vs 2.0679±0.4370 x10-7 cm·s-1), and catechins EC、EGC and ECG indicated no influences on uptake or transport.(3)The impact of green tea extract and catechins on the uptake of rosuvastatin in OATP1B1-HEK293 T cells: The uptake of rosuvastatin in positive transfected HEK293 T cells was significantly higher than the negative controls(6.9882 ± 0.6158 vs 2.4471 ± 0.4124 pmol·mg-1 protein, p<0.05), indicating a successful expression of transport activity. Moreover, 1.25 μg·m L-1 of green tea extract reduced the rosuvastatin uptake of rosuvastatin( 6.9882 ± 0.6158 vs 2.4471 ± 0.4124 pmol ·mg-1 protein, p<0.05), EGCG reduced the uptake( 6.9882 ± 0.6158 vs 3.1875 ± 0.9679 pmol·mg-1 protein, p <0.05), and EC、EGC and ECG did not change the values of uptake in statistics.(4)The impact of EGCG on the pharmacokinetics of rosuvastatin in rats: After combinative administration of EGCG and rosuvastatin, AUC0-∞ of rosuvastatin increased by 64.24%±47.44%(77.99±22.52 vs 47.48±11.71 h·μg·L-1, p<0.05)and its Cmax increased by 41.95%±83.38%(19.40±11.39 vs 13.66 ±7.01 ng·m L-1,p>0.05).Conclusion: It was indicated that the exposure amount of rosuvastatin in rats could be enhanced by green tea extract, giving rise to the possible mechanism of addition of small intestinal absorption and inhibition of drug transport by OATP1B1 in liver which reduced the hepatic distribution of rosuvastatin. Moreover, EGCG contained in green tea was found to be the main ingredient to affect the PK parameters of rosuvastatin. As a result, when taking statins in clinic, it is suggested not to take green tea and other drinks or food containing EGCG together. |
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