蒋逸;马增春;黄先菊;尤青;谭洪玲;王宇光;梁乾德;汤响林;肖成荣;高月;
JIANG Yi;MA Zeng-chun;HUANG Xian-ju;YOU Qing;TAN Hong-ling;WANG Yu-guang;LIANG Qian-de;TANG Xiang-lin;XIAO Cheng-rong;GAO Yue;College of Pharmacy,South-Central University for Nationalities;Institute of Radiation Medicine,Academy of Military Medical Sciences;

• 1:中南民族大学药学院
• 2:军事医学科学院放射与辐射医学研究所

摘要(Abstract)：

研究四物汤对Caco-2细胞P-糖蛋白(P-gp)功能和表达的影响。采用实时定量聚合酶链式反应(Q-PCR)分析Caco-2细胞上MDR1基因mRNA的表达量;采用流式细胞仪检测经四物汤水煎液处理后Caco-2细胞对罗丹明123摄取量的影响,以评价P-gp的外排功能;采用Western blotting法检测四物汤水煎液对Caco-2细胞P-gp蛋白表达量的影响。与空白对照组比较,四物汤水煎液3.3,5.0,10.0 g·L-1孵育Caco-2细胞24,48,72 h对MDR1基因mRNA的表达量具有上调作用,表明四物汤水煎液对MDR1具有诱导作用;四物汤水煎液作用于Caco-2细胞48 h后,细胞内罗丹明123的摄取量分别减弱了16.6%,22.1%(P<0.05),45.4%(P<0.01),提示长期应用四物汤水煎液,可增强Caco-2细胞P-gp的外排功能;四物汤水煎液孵育Caco-2细胞48 h后,上调了Caco-2细胞膜上P-gp蛋白表达量,表明四物汤水煎液对P-gp的蛋白表达量具有诱导作用。
To study the effect of Siwu decoction on the function and expression of P-glycoprotein( P-gp) in Caco-2 cells. The Realtime quantitative poly-merase chain reaction( Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells,in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2cells. Compared with the blank control group,after Caco-2 incubation with Siwu decoction at concentrations of 3. 3,5. 0,10. 0 g·L- 1for 24,48,72 h,the mRNA expression of MDR1 was up-regulated,suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h,the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16. 6%,22. 1%( P < 0. 05) and 45. 4%( P < 0. 01),indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h,the Pgp protein expression on Caco-2 cell emebranes,demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.

关键词(KeyWords)： 四物汤;P-gp;MDR1;Caco-2细胞;罗丹明123
Siwu decoction;P-gp;MDR1;Caco-2;Rhodamin 123

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金重点项目(81130067);国家自然科学基金面上项目(81073028,81274127)

作者(Authors): 蒋逸;马增春;黄先菊;尤青;谭洪玲;王宇光;梁乾德;汤响林;肖成荣;高月;
JIANG Yi;MA Zeng-chun;HUANG Xian-ju;YOU Qing;TAN Hong-ling;WANG Yu-guang;LIANG Qian-de;TANG Xiang-lin;XIAO Cheng-rong;GAO Yue;College of Pharmacy,South-Central University for Nationalities;Institute of Radiation Medicine,Academy of Military Medical Sciences;

参考文献(References)：

• [1]Lin J H.Drug-drug interaction mediated by inhibition and induction of P-glycoprotein[J].Adv Drug Deliv Rev,2003,55(1):53.
• [2]戴立波,方平飞,李焕德.P-gp调节对药物代谢动力学及药效学的影响[J].中南药学,2011,9(8):612.
• [3]吴芸,陈志鹏,蔡宝昌.桃红四物汤化学成分及药理作用的研究进展[J].中成药,2011,11(33):1965.
• [4]刘屏,宋森林,李宾,等.海洛因依赖者T细胞亚群变化及人参四物汤作用研究[J].中国药物依耐性杂志,2000,9(2):115.
• [5]张红丽,刘巧芬,章丽霞.黄芪四物汤结合西药及激光治疗糖尿病视网膜病变疗效观察[J].浙江中医药杂志,2010,11(45):804.
• [6]路晓钦,马增春,高月.四物汤药理研究进展[J].中国实验方剂学杂志,2001,4(7):56.
• [7]Zhong Qiu Liu,Zhi Hong Jiang,Liang Liu,et al.Mechanisms responsible for poor oral bioavailability of paeoniflorin:role of intestinal disposition and interactions with sinomenine[J].Pharmac Res,2006,12(23):2768.
• [8]杨雯,周惠芬,杨洁红,等.川芎嗪在Caco-2细胞单层模型的转运特征及对P-糖蛋白表达的影响[J].中草药,2013,44(5):581.
• [9]宋娟,唐靖,何娟,等.川芎嗪对Caco-2细胞p-糖蛋白功能和表达的影响[J].中南药学,2007,5(5):440.
• [10]高利臣.阿魏酸对药物代谢酶、转运体基因的影响及其与CYP2B6,POR和PXR基因多态性关系[D].长沙:中南大学,2012:29.
• [11]梁淼,马增春,肖成荣,等.四物汤及其配伍对大鼠肝脏P450酶活性及mRNA表达的影响[J].中国中药杂志,2013,38(21):3720.
• [12]路晓钦,高月,梁乾德,等.四物汤中不同组分对血虚模型小鼠造血功能的影响[J].中成药,2005,10(27):1189.
• [13]梁乾德,朱力军,谭洪玲,等.四物汤中糖类物质对辐射小鼠造血功能的影响[J].中草药,2006,37(2):226.
• [14]马增春,高月,谭洪玲,等.用分子中药组学技术研究四物汤补血的作用机理[J].世界科学技术——中医药现代化,2005,7(3):8.
• [15]何丹,颜苗,李焕德,等.甘草提取物及其主要成分对Caco-2细胞膜上P-gp功能和表达的影响[J].中国药学杂志,2010,45(10):571.
• [16]梁乾德,马百平,李卫华,等.用色谱和ESI 2 MS方法考察四物汤提取部位的化学成分[J].中国中药杂志,2004,29(4):334.