The Mechanism Of The Effect Of Breviscapine On Pharmacokinetics Of Atorvastain Based On CYP3A4 Enzyme

Purpose:The aim of this study was to investigate the effect of the breviscapine on pharmacokinetics of atorvastatin,and the effect of combined two drugs on the activity of CYP3A4 enzyme in rats.The changes in the protein expression and gene expression of the CYP3A4 enzyme after the breviscapine combined of atorvastatin from protein level and molecular biology level.To explore the mechanism that was the effects breviscapine on pharmacokinetics of atorvastatin.To provide reliable theoretical support for the combination of two drugs was synergistic clinical applications.To make the clinical application was more safely and effectively.Methods:Rats were randomly divided into,atorvastatin administration group,atorvastatin combined breviscapine administration group.The concentration of atorvastatin in rat plasma was determined by HPLC method,draw blood concentratio-time curve.The pharmacokinetic parameter was calculated with application software named DAS2.0.Rats were randomly divided into saline group,atorvastatin group,breviscapine group,breviscapine combined with atorvastatin group,and each group was administrated for 7 days continuously.First,the solution containing the specific probe substrate,midazolam,was prepared and incubated in the liver microsomes.RT-HPLC method was established to determine the concentration of the probe substrate in the liver microsomal incubation system.The effect of two drugs on the activity of CYP3A4 enzyme was evaluated by the concentration of the probe substrate.The gene expression of CYP3A4 was determined by using RT-PCR technology.The protein expression of CYP3A4 was determined by using Western Blot technology to explore the effects of the expression of CYP3A4 enzyme in each group.Results:(1)Compared with atorvastatin group,the pharmacokinetic parameters of atorvastatin increased in rats after atorvastatin combined with breviscapine,significantly reduced clearance,significantly prolonged the mean residence time,significantly increased in area under the curve(AUC).(2)Compared with the normal saline control group,there was no significant change in the activity of CYP3A4 enzyme in the atorvastatin group.Compared with the normal saline control group,there was significant change in the activity of CYP3A4 enzyme in the breviscapine group and breviscapine combined with atorvastatin drug group.In which the atorvastatin combined with breviscapine group had a very significant inhibitory effect on the activity of CYP3A4 enzyme.(3)Compared with the normal saline control group,there was no significant change in the expression of CYP3A4 enzyme gene in the atorvastatin group.The expression of CYP3A4mRNA in the group of breviscapine and breviscapine combined with the atorvastatin was significantly lower than that of the saline group.(4)The results of CYP3A4 enzyme protein expression show that compared with the normal saline control group,there was no significant change in the expression of CYP3A4 enzyme protein in the atorvastatin group.The expression of CYP3A4 protein in the group of atorvastatin combined with breviscapine and breviscapine group was significantly lower than that the saline group.Conclusions:After the combination of atorvastatin and breviscapine,breviscapine reduced the activity of the rat liver microsomal CYP3A4 enzyme,and reduce the expression of CYP3A4 gene and protein.The metabolic degree of atorvastatin of the substrate of CYP3A4 enzyme is slowed down in rats,significantly reduced clearance,significantly prolonged the mean residence time,significantly increased in area under the curve(AUC).Therefore,when atorvastatin and breviscapine are used together,it is possible to inhibit the activity of CYP3A4 enzyme of atorvastatin by inhibiting its gene transcription and protein translation level.The pharmacokinetic process of rats was changed,the metabolism of atorvastatin was slowed down and the curative effect was improved.