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

作　　者：蒋逸[1,2] 马增春[2] 黄先菊[1] 尤青[2] 谭洪玲[2] 王宇光[2] 梁乾德[2] 汤响林[2] 肖成荣[2] 高月[2] 

机构地区：[1]中南民族大学药学院,湖北武汉430074 [2]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《中国中药杂志》2015年第5期933-937,共5页China Journal of Chinese Materia Medica

基　　金：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)

摘　　要：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 Real- time 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-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5. 0, 10. 0 g ~ L- for 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 de- creased by respectively 16. 6%, 22. 1% （P 〈0. 05） and 45.4% （P 〈0. 01 ）, indicating that the long-term administration of Siwu de- coction 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 P- gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.

关 键 词：四物汤 P-GP MDR1 CACO-2细胞 罗丹明123 

分 类 号：R285[医药卫生—中药学]