中国中药杂志

2015, v.40(05) 933-937

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四物汤对Caco-2细胞P-糖蛋白功能和表达的影响
Effect of Siwu decoction on function and expression of P-glycoprotein in Caco-2 cells

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;

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;

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