四物汤对Caco—2细胞P—糖蛋白功能和表达的影响(1)
[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。
, http://www.100md.com
[关键词] 四物汤;P-gp;MDR1;Caco-2细胞;罗丹明123
[收稿日期] 2014-06-24
[基金项目] 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)
[通信作者] 高月，博士，研究员，Tel：(010)66931312，E-mail：gaoyue@nic.bmi.ca.cn;黄先菊，博士后，教授，Tel：(010)66930267，E-mail： huangxianju1972@yahoo.cn
[作者简介] 蒋逸，硕士研究生，Tel：(010)66930267，E-mail： jiangyizy@126.com
Effect of Siwu decoction on function and expression of
, 百拇医药
P-glycoprotein in Caco-2 cells
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
(1.College of Pharmacy， South-Central University for Nationalities， Wuhan 430074， China;
2. Institute of Radiation Medicine， Academy of Military Medical Sciences， Beijing 100850， China)
[Abstract] 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-1 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 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 P-gp protein expression on Caco-2 cell emebranes， demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp., 百拇医药(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)
, http://www.100md.com
[关键词] 四物汤;P-gp;MDR1;Caco-2细胞;罗丹明123
[收稿日期] 2014-06-24
[基金项目] 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)
[通信作者] 高月，博士，研究员，Tel：(010)66931312，E-mail：gaoyue@nic.bmi.ca.cn;黄先菊，博士后，教授，Tel：(010)66930267，E-mail： huangxianju1972@yahoo.cn
[作者简介] 蒋逸，硕士研究生，Tel：(010)66930267，E-mail： jiangyizy@126.com
Effect of Siwu decoction on function and expression of
, 百拇医药
P-glycoprotein in Caco-2 cells
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
(1.College of Pharmacy， South-Central University for Nationalities， Wuhan 430074， China;
2. Institute of Radiation Medicine， Academy of Military Medical Sciences， Beijing 100850， China)
[Abstract] 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-1 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 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 P-gp protein expression on Caco-2 cell emebranes， demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp., 百拇医药(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)