UPLC-MS/MS法研究黄芪甲苷对大鼠体内阿德福韦毒代动力学及肾转运的影响  

Effect of astragaloside on toxicokinetics and renal transport of adefovir in rats based on UPLC-MS/MS method

作　　者：郑圆 刘俊瑾 许丁 邵云云[2] 张晓 常壮鹏 侯锐钢[2] 贾晋生[3] ZHENG Yuan;LIU Jun-jin;XU Ding;SHAO Yun-yun;ZHANG Xiao;CHANG Zhuang-peng;HOU Rui-gang;JIA Jin-sheng(College of Pharmacy,Shanxi Medical University,Taiyuan 030001,China;Department of Pharmacy,the Second Hospital of Shanxi Medical University,Taiyuan 030000,China;Department of Pharmacy,Jincheng Coal Group General Hospital Affiliated to Shanxi Medical University,Jincheng 048006,China)

机构地区：[1]山西医科大学药学院,山西太原030001 [2]山西医科大学第二医院药剂科,山西太原030000 [3]山西医科大学附属晋煤集团总医院药剂科,山西晋城048006

出　　处：《中国药理学与毒理学杂志》2021年第4期297-304,共8页Chinese Journal of Pharmacology and Toxicology

基　　金：山西省自然科学基金(201901D111389);山西医科大学第二医院青年基金(201802-1);山西医科大学第二医院青年基金(201902-2)。

摘　　要：目的研究黄芪甲苷对大鼠体内阿德福韦毒代动力学的影响,并初步探究其降低阿德福韦肾蓄积的分子机制。方法(1)毒代动力学实验:12只SD雄性大鼠随机分为阿德福韦组(阿德福韦酯30 mg·kg^(-1))和黄芪甲苷干预组(阿德福韦酯30 mg·kg^(-1)+黄芪甲苷30 mg·kg^(-1)),每组6只,于ig给药后0,1,1.5,2,3,4,6,8,12和24 h经大鼠眼眶静脉丛取血,建立超高效液相色谱-串联质谱(UPLC-MS/MS)方法测定血浆阿德福韦浓度,进行方法学验证并计算毒代动力学参数;(2)肾转运实验:18只SD雄性大鼠随机分为生理盐水对照组、阿德福韦组和黄芪甲苷干预组,每组6只,盐水对照组和阿德福韦组ig给予生理盐水,黄芪甲苷组ig给予黄芪甲苷(30 mg·kg^(-1)),连续35 d,第8天时,除生理盐水对照组ig给予生理盐水外,其余各组ig给予阿德福韦酯30 mg·kg^(-1),连续28 d,应用实时荧光定量PCR测定大鼠肾组织有机阴离子转运体(Oat)和多药耐药转运体(Mrp)mRNA表达水平。结果(1)毒代动力学实验:建立了测定大鼠体内阿德福韦血药浓度的UPLC-MS/MS方法,该方法专属性强、灵敏度高且准确性好。与阿德福韦组相比,黄芪甲苷干预组大鼠血浆中阿德福韦药峰浓度(Cmax)和药时曲线下面积(AUC_(0-t)和AUC_(0-∞))显著降低(P<0.01),表观分布容积(V_(d))和血浆清除率(Cl)显著增加(P<0.01)。(2)肾转运实验:黄芪甲苷可显著降低肾中阿德福韦的浓度(P<0.01)。与生理盐水组相比,阿德福韦组大鼠肾组织中Oat1和Oat3 mRNA表达明显升高(P<0.01);黄芪甲苷干预后,与阿德福韦组相比,大鼠肾组织中Oat3 mRNA表达明显降低(P<0.01),Mrp4 mRNA表达明显升高(P<0.01)。结论黄芪甲苷可改变阿德福韦的毒代动力学参数,降低阿德福韦的肾蓄积,作用机制可能与肾小管上皮细胞Oat3和Mrp4 mRNA表达有关。OBJECTIVE To study the effect of astragaloside on the toxicokinetics of adefovir in rats and to investigate the molecular mechanism by which astragaloside reduces the renal accumulation induced by adefovir.METHORDS Toxicokinetic study:12 male SD rats were randomly divided into the adefovir group(adefovir dipivoxil 30 mg·kg^(-1))and astragaloside intervention group(adefovir dipivoxil30 mg·kg^(-1)+astragaloside 30 mg·kg^(-1)),with 6 rats in each.Blood samples were collected from the orbital venous plexus of rats at 0,1,1.5,2,3,4,6,8,12 and 24 h after ig administration.An ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for the determination of adefovir concentration in rat plasma was established,and validated before the toxicokinetic parameters were calculated.Renal transport study:18 SD male rats were randomly divided into the normal group,adefovir group and astragaloside intervention group,with 6 rats in each.The rats of the normal group and adefovir group were administrated intragastrically with normal saline,while those of the astragaloside intervention group with astragaloside(30 mg·kg^(-1))once a day for consecutive 35 days.On the 8 thday,the rats of the normal group were administrated intragastrically with normal saline,while those of other groups with adefovir dipivoxil(30 mg·kg^(-1))once a day for consecutive 28 days.The mRNA expression levels of organic anion transporters(Oats)and multidrug resistance transporters(Mrps)in rat renal tissue were determined by quantitative real-time fluorescence PCR technology.RESULTS Toxicokinetic study:the UPLC-MS/MS method established for the determination of adefovir concentration in rat plasma had strong specificity,high sensitivity and good accuracy.Cmax,AUC_(0-t) and AUC_(0-∞)in the astragaloside intervention group were significantly lower(P<0.01)than those in the adefovir group,whereas V_(d)and Cl were obviously higher(P<0.01).Renal transport study:astragaloside significantly reduced the concentration of adefovir in the kidney

关 键 词：超高效液相色谱-串联质谱法 阿德福韦 黄芪甲苷 毒代动力学 肾蓄积 

分 类 号：R285.5[医药卫生—中药学] R969[医药卫生—中医学]