板蓝根及所含靛蓝和靛玉红强烈抑制小鼠肾主要有机阴离子转运体Oat1,Oat2和Oat3  

Inhibition of Radix Isatidis and its constituents indigo and indirubin on major organic anion transporters Oat1,Oat2 and Oat3 in mouse kidneys

作　　者：奇锦峰[1] 孙晨[1] 王永辉 余文浩[1] 韩坚[1] 林梅[3] 张娜[1] 

机构地区：[1]广州中医药大学中药学院药理学教研室,广东广州510006 [2]河南省驻马店市人民医院药剂科,河南驻马店463000 [3]广州市佛山南海中医院西药房,广东广州528200

出　　处：《中国药理学与毒理学杂志》2014年第6期878-886,共9页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的探讨板蓝根颗粒剂(GRI)和饮片水煎剂(DRI)及其所含主要成分靛蓝和靛玉红对小鼠肾有机阴离子转运体(OAT)中3个主要亚型Oat1,Oat2和Oat3的影响。方法 NIH小鼠分别ig给予GRI0.615和2.460 g·kg-1,DRI 1.6和6.4 g·kg-1(生药量),靛蓝0.008和0.640 mg·kg-1,靛玉红0.0192和1.5360 mg·kg-1,每组60只(雌雄对半),每天2次,连续5 d。同时设丙磺舒(0.05 g·kg-1)阳性对照组和两种溶媒〔纯水和0.5%羧甲纤维素钠(CMC-Na)水溶液〕对照组及糊精加蔗糖(各1.5 g·kg-1)添加剂组。最后1次给予供试物后实施对-氨基马尿酸(PAH,iv,0.03 g·kg-1)清除实验,即在iv PAH后1.0,2.5,5.0,7.5,10.0和20.0 min时每组分别各取10只小鼠(雌雄对半),安乐处死收集全血制备血清,并迅速摘取双肾,右肾进行组织匀浆后测定PAH蓄积量,左肾组织用于提取总m RNA。每组另取10只小鼠(雌雄各半),同样给药处理,按Nakakariya法做肾切片进行摄取PAH实验。用Kiguchi法测定血清和肾组织匀浆液中PAH浓度。以药动学软件(DAS 2.0)计算血清及肾组织中PAH的主要药动学参数。以实时定量PCR法测定小鼠肾组织Oat1,Oat2及Oat3 m RNA表达。结果与纯水对照组比较,0.5%CMC-Na对照组各项检测指标均无显著差异。与两种溶媒对照组相比,GRI 2.460 g·kg-1,靛蓝0.640 mg·kg-1和靛玉红1.5360 mg·kg-1组消除半衰期(t1/2β)显著延长(P<0.05);各供试物组分布容积(Vd)和清除率(Cl)均显著减少(P<0.01),曲线下面积(AUC0→20 min)均显著增加(P<0.01);由采血时间段内各组肾组织中的PAH蓄积量所求得的AUC0→20 min显著大于同期对照组(P<0.05,P<0.01),且肾AUC0→20 min与血液AUC0→20 min的比值在各组间无显著差异。各剂量供试物均可使肾切片摄取PAH的量显著少于对照组(P<0.05,P<0.01)。与纯水/CMC对照组相比,GRI 2.460 g·kg-1,DRI 6.4 g·kg-1,靛蓝0.640 mg·kg-1,靛玉红1.5360 mg·kg-1组小鼠肾组织Oat1,Oat2及Oat3 m RNA表达除靛玉红组Oat2 m RNA(P<0.05)、OBJECTlVE To investigate the inhibition of Radix lsatidis and its major constituents indigo and indirubin on three principal subtypes of organic anion transporters （ OATs） , Oat1, Oat2 and Oat3 in vivo in mice. METHODS Granules of Radix lsatidis （ GRl） 0.615 and 2.46 g·kg^-1 , decoction of Radix lsatidis （ DRl） 1.6 and 6.4 g·kg^-1 , indigo 0.008 and 0.64 mg·kg^-1 and indirubin 0.0192 and 1.536 mg·kg^-1 were ig given to the NlH mice （60 mice per group）, twice a day, for 5 d while four control groups were set up, including vehicle of water, 0.5%sodium carboxymethyl cellulose （ CMC） , positive control probe-necid （0.05 g·kg^-1） and additives of sucrose plus dextrin （1.5 g·kg^-1 each） groups. After the last dosing of the test samples, para-aminohippuric acid （ PAH） clearance test was conducted. All the mice were iv given PAH 0.03 g·kg^-1 and 1, 2.5, 5, 7.5, 10 and 20 min later before 10 mice per group were euthanized to collect whole blood and the kidneys were quickly removed. Each right kidney was homoge-nized to analyze the PAH accumulations and each left kidney to extract total mRNA for analysis of Oat1, Oat2 and Oat3 gene expressions using quantitative real-time PCR. The concentrations of PAH in sera and in kidney homogenates were determined by the method of Kiguchi. Major pharmacokinetic parame-ters of PAH in sera were calculated by pharmacokinetic software （ DAS2.0） . PAH uptake test for kidney slices was performed on another group of NlH mice according to the method of Nakakariya. RESULTS There was no significant difference between water control group and 0.5%CMC group in all the examined items. Compared with the vehicle control groups （ water and 0. 5%CMC group ） , elimination half time （ t1/2β） of PAH in GRl 2.46 g·kg^-1 ,indigo 0.64 mg·kg^-1 and indirubin 1.536 mg·kg^-1 groups was signifi-cantly prolonged （P〈0.05）, the total clearance （Cl） and volume of distribution （Vd） were obviously reduced （ P〈0.01） and the area under the curve �

关 键 词：板蓝根 靛蓝 靛玉红 对-氨基马尿酸 有机阴离子转运体 

分 类 号：R285.5[医药卫生—中药学]