目的：探讨板蓝根颗粒剂( GRl)和饮片水煎剂( DRl)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 NlH小鼠分别ig给予GRl 0.615和2.460 g·kg－1,DRl 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mRNA。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 mRNA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, GRl 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,GRl 2.460 g·kg－1,DRl 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 mRNA表达除靛玉红组Oat2 mRNA( P<0.05)、GRl组Oat3 mRNA( P<0.01)表达水平被显著上调外,其余均被显著下调( P<0.05,P<0.01)。结论 GRl、DRl、靛蓝、靛玉红在所用剂量下对小鼠肾组织Oat1,Oat2及Oat3均有明显抑制作用,GRl和DRl的这种抑制作用可能主要来自其所含的靛蓝和靛玉红成分。
目的：探討闆藍根顆粒劑( GRl)和飲片水煎劑( DRl)及其所含主要成分靛藍和靛玉紅對小鼠腎有機陰離子轉運體( OAT)中3箇主要亞型Oat1,Oat2和Oat3的影響。方法 NlH小鼠分彆ig給予GRl 0.615和2.460 g·kg－1,DRl 1.6和6.4 g·kg－1(生藥量),靛藍0.008和0.640 mg·kg－1,靛玉紅0.0192和1.5360 mg·kg－1,每組60隻(雌雄對半),每天2次,連續5 d。同時設丙磺舒(0.05 g·kg－1)暘性對照組和兩種溶媒〔純水和0.5％羧甲纖維素鈉( CMC-Na)水溶液〕對照組及糊精加蔗糖(各1.5 g·kg－1)添加劑組。最後1次給予供試物後實施對-氨基馬尿痠( PAH, iv,0.03 g·kg－1)清除實驗,即在iv PAH後1.0,2.5,5.0,7.5,10.0和20.0 min 時每組分彆各取10隻小鼠(雌雄對半),安樂處死收集全血製備血清,併迅速摘取雙腎,右腎進行組織勻漿後測定PAH蓄積量,左腎組織用于提取總mRNA。每組另取10隻小鼠(雌雄各半),同樣給藥處理,按Nakakariya法做腎切片進行攝取PAH實驗。用Kiguchi法測定血清和腎組織勻漿液中PAH濃度。以藥動學軟件( DAS 2.0)計算血清及腎組織中PAH的主要藥動學參數。以實時定量PCR法測定小鼠腎組織Oat1,Oat2及Oat3 mRNA錶達。結果與純水對照組比較,0.5％CMC-Na對照組各項檢測指標均無顯著差異。與兩種溶媒對照組相比, GRl 2.460 g·kg－1,靛藍0.640 mg·kg－1和靛玉紅1.5360 mg·kg－1組消除半衰期( t1/2β)顯著延長( P<0.05)；各供試物組分佈容積( Vd )和清除率( Cl)均顯著減少(P<0.01),麯線下麵積(AUC0→20 min)均顯著增加(P<0.01)；由採血時間段內各組腎組織中的PAH蓄積量所求得的AUC0→20 min顯著大于同期對照組( P<0.05,P<0.01),且腎AUC0→20 min與血液AUC0→20 min的比值在各組間無顯著差異。各劑量供試物均可使腎切片攝取PAH的量顯著少于對照組( P<0.05,P<0.01)。與純水/CMC對照組相比,GRl 2.460 g·kg－1,DRl 6.4 g·kg－1,靛藍0.640 mg·kg－1,靛玉紅1.5360 mg·kg－1組小鼠腎組織Oat1,Oat2及Oat3 mRNA錶達除靛玉紅組Oat2 mRNA( P<0.05)、GRl組Oat3 mRNA( P<0.01)錶達水平被顯著上調外,其餘均被顯著下調( P<0.05,P<0.01)。結論 GRl、DRl、靛藍、靛玉紅在所用劑量下對小鼠腎組織Oat1,Oat2及Oat3均有明顯抑製作用,GRl和DRl的這種抑製作用可能主要來自其所含的靛藍和靛玉紅成分。
목적：탐토판람근과립제( GRl)화음편수전제( DRl)급기소함주요성분전람화전옥홍대소서신유궤음리자전운체( OAT)중3개주요아형Oat1,Oat2화Oat3적영향。방법 NlH소서분별ig급여GRl 0.615화2.460 g·kg－1,DRl 1.6화6.4 g·kg－1(생약량),전람0.008화0.640 mg·kg－1,전옥홍0.0192화1.5360 mg·kg－1,매조60지(자웅대반),매천2차,련속5 d。동시설병광서(0.05 g·kg－1)양성대조조화량충용매〔순수화0.5％최갑섬유소납( CMC-Na)수용액〕대조조급호정가자당(각1.5 g·kg－1)첨가제조。최후1차급여공시물후실시대-안기마뇨산( PAH, iv,0.03 g·kg－1)청제실험,즉재iv PAH후1.0,2.5,5.0,7.5,10.0화20.0 min 시매조분별각취10지소서(자웅대반),안악처사수집전혈제비혈청,병신속적취쌍신,우신진행조직균장후측정PAH축적량,좌신조직용우제취총mRNA。매조령취10지소서(자웅각반),동양급약처리,안Nakakariya법주신절편진행섭취PAH실험。용Kiguchi법측정혈청화신조직균장액중PAH농도。이약동학연건( DAS 2.0)계산혈청급신조직중PAH적주요약동학삼수。이실시정량PCR법측정소서신조직Oat1,Oat2급Oat3 mRNA표체。결과여순수대조조비교,0.5％CMC-Na대조조각항검측지표균무현저차이。여량충용매대조조상비, GRl 2.460 g·kg－1,전람0.640 mg·kg－1화전옥홍1.5360 mg·kg－1조소제반쇠기( t1/2β)현저연장( P<0.05)；각공시물조분포용적( Vd )화청제솔( Cl)균현저감소(P<0.01),곡선하면적(AUC0→20 min)균현저증가(P<0.01)；유채혈시간단내각조신조직중적PAH축적량소구득적AUC0→20 min현저대우동기대조조( P<0.05,P<0.01),차신AUC0→20 min여혈액AUC0→20 min적비치재각조간무현저차이。각제량공시물균가사신절편섭취PAH적량현저소우대조조( P<0.05,P<0.01)。여순수/CMC대조조상비,GRl 2.460 g·kg－1,DRl 6.4 g·kg－1,전람0.640 mg·kg－1,전옥홍1.5360 mg·kg－1조소서신조직Oat1,Oat2급Oat3 mRNA표체제전옥홍조Oat2 mRNA( P<0.05)、GRl조Oat3 mRNA( P<0.01)표체수평피현저상조외,기여균피현저하조( P<0.05,P<0.01)。결론 GRl、DRl、전람、전옥홍재소용제량하대소서신조직Oat1,Oat2급Oat3균유명현억제작용,GRl화DRl적저충억제작용가능주요래자기소함적전람화전옥홍성분。
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 ( AUC0-20 min ) of PAH in all the tested groups was signifi-cantly increased ( P<0.01) . AUC0-20 min obtained from renal PAH accumulations within the checked time was significantly higher ( P<0.05, P<0.01) than in the vehicle control group. But there was in no signifi-cant difference between all the study groups in kidney-to-plasma AUC ratios. PAH uptake results by kidney slices were significantly lower ( P<0. 05, P<0. 01 ) than in vehicle control group in every two dosages of all the four samples tested. Compared with vehicle control group, the mRNA expressions of Oat1, Oat2 and Oat3 were obviously ( P<0.05, P<0.01) and abnormally regulated in the groups of GRl 2.46 g·kg-1, DRl 6.4 g·kg-1, indigo 0.64 mg·kg-1 and indirubin 1.536 mg·kg-1. CONCLUSlON The renal Oat1, Oat2 and Oat3 of mice are significantly inhibited by GRl, DRl, indigo and indirubin. The inhibitory function of Radix lsatidis probably stems from indigo and indirubin contained in it.
