Effect Of QiShenYiQi Dropping Pills On The Pharmacokinetics Of Atorvastatin Based On Metabolic Enzymes And Transporters

Part one Effects of QiShenYiQi dropping pills on pharmacokinetics of atorvastatin in ratsObjective:To establish a method for the determination of atorvastatin（atorvastatin,ATV）and its metabolites o-hydroxy atorvastatin（ortho-hydroxy atorvastatin,o-ATV）and p-hydroxy atorvastatin（para-hydroxy atorvastatin,p-ATV）in rat plasma,and to investigate the effects of Qi Shen Yiqi dropping pills on the pharmacokinetics of atorvastatin after oral and intravenous injection in rats.Methods:20 male Sprague–Dawley rats were randomly divided into four groups:Control oral group（atorvastatin 10 mg/kg）,Treatment oral group（atorvastatin 10 mg/kg after QiShenYiQi dropping pills 675 mg/kg once a day for 7 days）,Control intravenous group（atorvastatin 2 mg/kg）,Treatment intravenous group（atorvastatin 2 mg/kg after QiShenYiQi dropping pills 675mg/kg once a day for 7 days）.Blood samples were collected at different time points and analyzed the blood concentration of atorvastatin,p-ATV and o-ATV by UPLC-MS/MS after liquid-liquid extraction.The chromatographic separation was carried out on a Waters XBridge BEH C18 column.For the detection,the mass spectrometer was set to negative ionization mode.The multiple reaction monitoring mode was used to quantify the analytes and IS.Results:The calibraton curve for ATV,o-ATV and p-ATV in rat plasma samples showed good linearity with the test concentration rang from 0.5-250ng/ml.Compared with control oral group,AUC_0-24h and C_max of ATV increased80%and 95%,t_1/2 and V decreased 74%and 80%,C_max of p-ATV increased154%,t_1/2 and V decreased 61%and 76%in treatment oral group.No change was observed in intravenous group.Conclusions:A sensitive and specific UPLC-MS/MS method to quantify ATV,o-ATV and p-ATV in rat plasma samples was established in the study.The method was applied for assessing the effect of QiShenYiQi dropping pills on pharmacokinetics of ATV,o-ATV and p-ATV.According the results,the plasma concentration of ATV and o-ATV was observed in oral group,while no change was observed in intravenous group,implying that QiShenYiQi dropping pills and atorvastatin may have interaction in gastrointestinal tract.Part two Effects of QiShenYiQi dropping pills on the expression of metabolic enzymes and transporters in ratsObjective:To investigate the rat Cyp3a1,Cyp3a2,Slco1b2 m RNA expression in liver and Abcb1a,Slco2b1,Abcg2,Abcc2 m RNA expression in jejunum and ileum,the study utilizes Real-time fluorescence quantitive PCR（q RT-PCR）.Methods:Rats was anesthetized with pentobarbital after plasma samples collected,and the tissues of liver,jejunum and jejunum were taken and preserved at-80℃.The experiment was carried out in ABI 7500 PCR.Rats m RNA was extracted using DP419 total RNA extraction kit,KR118m RNA reverse transcription kit,F205m RNA amplification kit,and internal reference sequence wasβ-actin.Pre-denaturation stage of amplification program was set at 95℃with 15 minutes.PCR reaction stage was set at 95℃with 30 seconds for denaturation,and 60℃with 32 seconds for annealing extension,repeated for 40 cycles.Results:Compared with control oral group,Abcb1a expression in jejunum decrased 67%and Abcc2 expression in ileum decrased 63%.While Abcb1a expression in jejunum decrased 41%and Abcc2 expression in ileum decrased 59%comparing control intravenous group.No significant change was detected in other m RNAs.Conclusions:No matter in oral or intravenous groups,only Abcb1a and Abcc2 was observed in jejunum and ileum separately.The results indicated QiShenYiQi dropping pills may down regulated the tansporters expression of Abcb1a in jejunum and Abcc2 in ileum,resulting in less ATV effluxion and more ATV exposure in rats.