Study On Liver Targeting Effect Of Vinegar Processing Radix Bupleuri On Oxymatrine In Mice

Speciality:PharmaceuticsAuthor:Youjun ChenTutor:Ruizhi ZhaoAIM:Study the influence of vinegar processing radix bupleuric(VPRB) on the distribution of Oxymatrine(OMT) and its main metabolite Matrine(MT),in rats body,and verify the liver target enhancing effect of VPRB on Matrine category alkaloids.Methods:1.Oxymatrine and Matrine concerntration of tisssues were determinated by High-performance liquid chromatography mass spectrometry.The chromatographic column is Interlsil ODS-SP(5μm 2.1X150mm),the mobile phase consisted of methanol-10mmol ? L^-1 NH₄Ac(adjust the pH value to 3.5 by acetic acid),flow rate is 300μL ? min(-1),column temperature is room temperature,the injection volumn is 10μL.The tissues were homogenated with 10 times water,and the homogenate and plasma samples were extracted by trichlormethane with 2%n-butanol through liquid-liquid extraction.Oxymatrine(m/z265.3→247.2),Matrine(m/z 249.5→148.2) and internal standard Finasteride(m/z373.5→305.6) were detected by the positive electrospray ionization(ESI)-MS method under multiple reaction monitoring(MRM) mode.2.The influence of VPRB on the distribution of OMT and MT in rat body was evaluated by vivo distribution experiment.336 male rats were divided into two big groups according the dose of oxymatrine,oxymatrine lower and higher group(10 and 80mg·kg(-1)).At each big group,oxymatrine group as control and oxymatrine coadministered with three different doses of VPRB(400,800,1200 mg·kg(-1)) intragastric administration.Samples were collected at the time points of 0.167,0.5,1,2,4,8 hours for oxymatrine lower group,and 0.167,0.333, 0.667,1,1.5,2,4,8 hours for oxymatrine higher group.The samples were detected by HPLC-MS/MS and analysis the pharmacokinetic process.The influences of VPRB on the distribution of OMT and MT were evaluated through the pharmacokinetic parameters,such as Tmax,Cmax,AUC,AUC_OMT/AUC_MT and AUCsum.3.The target effect of different tissues of VPRB on OMT and MT was evaluated through the parameters of Relative uptake efficiency(Re) and Relative targeting efficiency(RTE).Results:1.The linearity of oxymatrine and matrine both ranged from 4 to 2000 ng ? ml(-1), the detection limits of them was 4ng ? mL(-1).The recovery of this method ranged from 87.1-110.3%.The values of RSD of within day and between day were less than 14%.The recovery rates of extraction were above 58.6%.2.Oxymatrine and Matrine processing in plasma,liver,heart,spleen,lung and kidney for OMT higher dosage group fit an open two-compartment model,but in most tissues,they fit a one-cmpartment model for OMT lower dosage group.VPRB increased the Cmax and AUC of both OMT and MT in liver.In OMT higher group,OMT Cmax of VPRB-M group and VPRB-H group increased by above 80 percent as compared with control group respectively,and the Cmax of MT in both groups were also increased.The AUC of OMT in VPRB-M group and H group were 2.55 and 2.53 times larger compared to control group,and the MT AUC of these two groups increased 12.09%and 16.36%respectively.In OMT lower group,the AUC of OMT in VPRB-L group and VPRB-H group are larger than the control group,the increased value of VPRB-H group is 31.82%, and meanwhile the Cmax and AUC of plasma,heart lung and kidney were tending to decline.In OMT higher group,compared to the control group,VPRB-M group and VPRB-H group increased the Re and RTE of liver OMT,the Re of VPRB-M and H group were 2.55 and 2.53 respectively,and the Re of other tissues were all less than 1.In all VPRB groups the liver RTE were positive value,the OMT RTE of VPRB-L, M,H groups were 0.40,2.16 and 2.11 respectively,and those of MT were 0.13, 0.21and 0.33 respectively.In Oxymatrine lower group,the liver OMT RTE of VPRB-L,M and H groups were 0.04,0.10 and 0.33 respectively,and the RTE of MT were 0.29,0.1 and 0.49 respectively.VPRB high dose had the strongest effect in both OMT higher group and lower group.Conclosion: HPLC-MS/MS method is accurate,highly sensitive,simple and convenient, time saving for the simultaneous determination of oxymatrine and matrine concentrations in tissues and plasma of rat.The targeting enforcing effect of VPRB was related with the dose of VPRB, high and medium dose of VPRB could enhance the targeting efficiency of OMT and MT significantly.VPRB focus the effect of other drug on liver by increasing the uptake of liver and meanwhile decreasing the uptake of other tissues.The results of this paper imply that VBRB might be a potential drug for target therapy.