摘要
目的初步检测缩泉丸和真武汤对小鼠肾脏有机阴离子转运体（Oats）中的2个主要亚型（Oat1,Oat3）的影响。方法以缩泉丸和真武汤及方中各药水煎剂、丙磺舒、氢氯噻嗪给NIH小鼠灌胃,每天2次,连续5 d,另设纯水对照组。末次给药后所有受试鼠逐一经尾静脉注入对氨基马尿酸（para-aminohippuric acid,PAH）30mg·kg^-1,然后在1,2.5,5,7.5,10,15,20 min时分别从各组各取10只小鼠处死收集全血,并迅速摘取双肾,测定血清及肾匀浆液中的PAH浓度,以药动学软件（DAS2.0）求出血清及肾组织匀浆液中PAH的主要药动学参数：消除半衰期（t1/2β）,分布容积（Vd）,清除率（CL）和药-时曲线下面积（AUC0-20 min）;从各组右肾提取总mRNA,以实时荧光定量PCR法比较小鼠肾脏中Oat1、Oat3 mRNA的表达量。另从各组各取10只小鼠,末次给药后1 h,断头处死后,取右肾做肾切片摄取PAH试验。结果与纯水对照组比较,真武汤组、茯苓组t1/2β显著延长（P〈0.01）,缩泉丸、乌药、附子各组t1/2β显著缩短（P〈0.05,P〈0.01）;缩泉丸、益智仁、山药、真武汤、白芍、生姜、茯苓各组Vd显著降低（P〈0.05,P〈0.01）,附子组Vd升高（P〈0.01）;缩泉丸、山药、真武汤、白术、生姜、茯苓各组CL均显著降低（P〈0.05,P〈0.01）;除益智仁、白芍、茯苓各组AUC0-20 min无显著变化外（P〉0.05）,其他供试物组均显著增大（P〈0.01）。各供试物组肾脏与血清AUC0-20 min比值均在0.247~0.475之间,与纯水对照组相似;肾切片摄取PAH试验显示,与纯水对照组比较,缩泉丸、乌药、山药、真武汤、白术、白芍、附子、生姜、茯苓各组摄取PAH量明显降低（P〈0.05,P〈0.01）。与纯水对照组比较,益智仁组、附子组小鼠肾脏Oat1 mRNA表达水平显著上调（P〈0.01）,而缩泉丸组、真武汤组Oat1 mRNA表达水平则显著下调（P〈0.01）;缩泉丸、益智仁、附子各组小鼠肾脏Oat3 mRNA表达水平显著上调（P〈0.01）,而真武汤组Oat3 mRNA表达水平则显著下调（P〈0.01）。结论缩泉丸及其君药益智仁、真武汤及其君药附子可能通过影响有机阴离子转运体的2个主要成员Oat1,Oat3的表达导致肾脏损伤。
Objective To investigate the effects of Suoquan Pills and Zhenwu Decoction on renal organic an ion transporter Oat1 and Oat3 in vivo in mice. Methods NIH mice were orally treated with water decoction of investigated samples two times a day for five continuous days. Mice in blank control group were given pure water orally. After last medication,all of the mice were i.v. given para-aminohippuric acid（PAH）at one dose of 30 mg·kg-1through the caudal vein,and 1,2.5,5,7.5,10,15,20 min later,10 mice from each group were executed for collecting whole blood at each time point respectively and kidneys were quickly removed. The left kidney was homogenized for PAH accumulation test（protein contents were quantified by Folin-phenol method）. The concentrations of PAH in serum and renal homogenate were determined. Pharmacokinetic analysis（t1/2β,CL,Vd,AUC0-20min） of PAH in blood and renal homogenate was performed by using pharmacokinetic Software（DAS2.0）,and the parameters covered elimination half life（t1/2β）, distribution volume（Vd）, total body clearance（CL） and concentration-time area under curve（AUC0-20min）. Total mRNA was extracted from the right kidney of mice,and the m RNA expression levels of renal Oat1,Oat3 were assayed by real-time fluorescent quantitation PCR. Another 10 mice from each group were executed one hour after last medication to obtain the right kidney for PAH uptake test. Results Compared with the blank control group,t1/2β was obviously prolonged in Zhenwu Decoction group and Poria（Fuling） group（P0.01）,and was shortened in Suoquan Pills group, Radix Linderae（Wuyao） group, and Radix Aconiti（Fuzi） group（P0.05,P0.01）. Vd was decreased in Suoquan Pills group,Fructus Alpiniae Oxyphyllae（Yizhiren）group,Fuling group,Zhenwu Decoction group,Radix Paeoniae Alba（Baishao）group,and Rhizoma Zingiberis Recens（Shengjiang） group（P0.05,P0.01）, and was increased in Fuzi group（P〈0.01）. CL in the medication groups of Suoquan pills, Shanyao,Zhenwu Decoction,Baizhu,Shenjiang and Fuling was decreased（P0.05,P0.01）. AUC0-20 minwas increased in the medication groups except for Yizhiren,Baishao,and Fuling（P0.01）. The ratio of renal AUC to serum AUC was in the range of 0.247~0.475,similar to that of blank control group. The results of PAH uptake test showed that PAH content was decreased in the medication groups of Suoquan pills,Wuyao,Shanyao,Zhenwu Decoction,Baizhu,Baishao, Fuzi, Shengjiang, Fuling（P〈0.05, P0.01）. Compared with blank control group, renal Oat1 mRNA expression level in Yizhiren group and Fuzi group was up-regulated（P0.01）,but was down-regulated in Suoquan Pills group and Zhenwu Decoction group（P〈0.01）. Renal Oat3 mRNA expression level was up-regulated in Suoquan Pills group,Yizhiren group and Fuzi group（P〈0.01）,but was down-regulated in Zhenwu Decoction group（P〈0.01）.Conclusion Suoquan Pills and their monarch drug Yizhiren, Zhenwu Decoction and its monarch drug Fuzi have an effect on renal injury through regulating the expression of renal organic anion transporter Oat1 and Oat3 in vivo in mice.
出处
《中药新药与临床药理》
CAS
CSCD
北大核心
2015年第3期342-347,410,共7页
Traditional Chinese Drug Research and Clinical Pharmacology
关键词
缩泉丸
真武汤
有机阴离子转运体
对氨基马尿酸
清除
MRNA表达
Suoquan Pills Zhenwu Decoction Organic anion transporter Para-aminohippuric acid Renal clearance test mRNA expression
