Effection Of Huangqishengmai Liquid On Three Isoforms Of P450 Cytochromes(CYP450) In Rats

Objectives Combined with the earlier studies in this group about the effect of active ingredients of Chinese medicine on Cytochrome P450(CYP450) activity in vivo and in vitro, selecting five components in Huangqishengmai liquid and researching the effect on CYP1A2, CYP2C9, CYP3A4 activity in rats liver in vivo and in vitro, expouding the effect rule of Huangqishengmai liquid on CYP450 and evaluating the safety of drug-drug interaction.Methods1 By the method of HPLC-DAD determined the content of Lobetyolin, Ruscogenin, Schizandrin B, Schisantherin A in Huangqishengmai liquid;2 By the method of “Cocktail” probe drugs detected the amount of metabolism of three specific probe drugs of CYP1A2, CYP2C9 and CYP3A4 in rat serum in every group, we have calculated pharmacokinetics parameters and drew the concentration-time curve by the DAS2.0 software, and comparing AUC(0-t);3 The Rat Liver Microsomes in vitro incubation combined with “Cocktail” probe substrates, study the influence of the active ingredients of Huangqishengmai liquid(astragaloside、Lobetyolin, Ruscogenin, Schizandrin B, Schisantherin A) and all components in it on three major CYPs(CYP1A2, CYP2C9 and CYP3A4) activity in RLM in vitro. We also had detected and compared the maximum reaction velocity of three specific probe drugs of CYP isoforms CYP1A2, CYP2C9 and CYP3A4, namely caffeine, tolbutamide and dapsone.Result1 The peak of four components in Huangqishengmai liquid, namely Lobetyolin, Ruscogenin, Schizandrin B, Schisantherin A, were separated by baseline, there no measurement interference, that is a simple and quick chromatography method has been established for simultaneously determinating content of multicomponent.2 The effect of drugs on CYP450 isoforms in “Cocktail” probe drug in vivo:(1) CYP1A2: Radix Ophiopogonis could induce the activity of CYP1A2 and fructus schizandrae could inhibit its activity;(2) CYP2C9: Astragalus membranaceus、Codonopsis pilosula and fructus schizandrae could increase the activity of CYP2C9;(3) CYP3A4: Huangqishengmai liquid and Radix Ophiopogonis could increase the activity of CYP3A4 but fructus schizandrae could decrease its activity;3 The effect of drugs on CYP450 isoforms in RLM in vitro(1) CYP1A2: Huangqishengmai liquid,astragaloside and Schizandrin B could inhibit the activity of CYP1A2;(2) CYP2C9: Lobetyolin, Schisantherin A could induce its activity;(3) CYP3A4: Huangqishengmai liquid, Schisantherin A and Ruscogenin could induce its activity, and Schizandrin B could inhibit its activity.Conclusions 1. In vitro, Compared to control, the V of astragaloside and Schizandrin B for CYP1A2 specific probe drug, namely caffeine, decreased 0.50 and 0.20, respectively. Therefore, we speculated the main sources of inhibition of Huangqishengmai liquid on CYP1A2 are astragaloside and Schizandrin B. Similarly, Ruscogenin is the main reasons for induction of Huangqishengmai liquid on CYP3A4, because of the V increasing 1.55 compared to control.2. Huangqishengmai liquid has induced CYP3A4 activity in vivo and in vivo. When Huangqishengmai liquid combinated with medicines which via the CYP3A4 metabolism, it could occur drug-drug interactions and produce adverse drug reactions.3 In vitro, compared to the blank group, the area of concentration-time curve of Radix Ophiopogonis for CYP3A4 specific probe drug, namely dapsone, decreased 0.29, respectively. Therefore, we considered Radix Ophiopogonis is the main causes for induction of Huangqishengmai liquid on CYP3A4.