异丹叶大黄素在Caco—2细胞模型上转运机制的研究(1)
[摘要]异丹叶大黄素具有抗炎、抗氧化、抗癌等药理活性。该文研究了异丹叶大黄素在Caco-2细胞模型上的吸收转运机制。采用UPLC法，应用PDA检测器在310 nm处对异丹叶大黄素进行含量测定并计算表观渗透系数Papp。考察不同浓度异丹叶大黄素在Caco-2细胞上的毒性以确定转运实验的给药浓度。探讨了时间、浓度、温度、转运体抑制剂对异丹叶大黄素在体外细胞模型上跨膜转运的影响。实验结果表明，异丹叶大黄素在10～60 μmol·L-1，孵育14 h内，对Caco-2细胞没有明显毒性。异丹叶大黄素在Caco-2细胞模型上的转运具有一定的浓度依赖性，其表观渗透系数Papp大于10×10-6 cm·s-1，易被Caco-2细胞吸收。 BL侧的转运量在3 h达到最大值，6 h有所下降。4 ℃条件下，Papp(AP-BL)与Papp(BL-AP) 均比37 ℃条件显著减小。加入P-gp转运体抑制剂维拉帕米后Papp(AP-BL)明显增大；加入MRP转运体抑制剂丙磺舒和MK-571后，Papp(BL-AP)明显减小。研究结果提示，异丹叶大黄素在Caco-2细胞模型上的转运方式主要是被动扩散，P-gp及MRP可能参与了异丹叶大黄素的外排转运。
, 百拇医药
[关键词]Caco-2细胞单层膜模型； 异丹叶大黄素； 转运机制
[Abstract]Isorhapontigenin (ISO) is suggested to have many different kinds of pharmacology activities， such as anti-inflammatory effect， anti-oxidation effect and anti-cancer effect. This paper mainly discussed the transport mechanism of ISO in Caco-2 cell models. The concentration of ISO was determined by UPLC method with PDA detector at 310 nm， and then the apparent permeability coefficient Papp was calculated. The cytotoxic of different concentrations of ISO was investigated on Caco-2 cells to determine the concentration of drug administration. The effects of ISO concentration， time， temperature and transporter inhibitors on the transport of ISO were investigated. The test results showed that， ISO didn′t have significant cytotoxicity at 10-60 μmol·L-1 in 14 hours. The transportation of ISO on Caco-2 cells was related to the concentration to a certain extent. Papp of ISO was higher than 10×10-6 cm·s-1 and ISO was absorbed easily by Caco-2 cells. The transport volume of ISO at BL side reached maximum at 3 h and was slightly decreased at 6 h. Papp (AP-BL) and Papp(BL-AP) at 4 ℃ were lower than those at 37 ℃. Papp (AP-BL) of ISO was significantly increased after adding P-gp inhibitor verapamil and Papp (BL-AP) of ISO was significantly decreased after adding MRP-2 inhibitor (probenecid or MK-571). The results suggested that transport mode of ISO was mainly passive diffusion in Caco-2 cell models， and P-gp and MRP may be involved in the transport of ISO.
, 百拇医药
[Key words]Caco-2 cell monolayer model； isorhapontigenin； transport mechanism
異丹叶大黄素(isorhapontigenin，ISO)存在于买麻藤类植物中，具有良好的抗炎[1-2]，抗氧化[3-5]，治疗胰岛素抵抗及肥胖[6]等活性。近年来，有研究发现其有显著的抗膀胱癌活性[7-11]。有研究表明，小鼠灌胃不同浓度的异丹叶大黄素，采用LC-MS/MS测定其不同时间点的血药浓度，发现异丹叶大黄素在小鼠体内迅速达峰，消除迅速，代谢较快[9]。Caco-2细胞来源于人体结肠腺癌细胞，经培养分化后可形成与人体小肠上皮细胞一致的形态及功能，含有广泛存在于人体小肠上皮细胞的药物代谢酶及各种转运体[12]，可以用来研究药物在体外吸收和代谢情况。该模型是目前应用最广泛，最经典的用于研究药物吸收转运特性的体外模型。, 百拇医药(袁紫朔 张婷婷 金波 李彤 马辰)
, 百拇医药
[关键词]Caco-2细胞单层膜模型； 异丹叶大黄素； 转运机制
[Abstract]Isorhapontigenin (ISO) is suggested to have many different kinds of pharmacology activities， such as anti-inflammatory effect， anti-oxidation effect and anti-cancer effect. This paper mainly discussed the transport mechanism of ISO in Caco-2 cell models. The concentration of ISO was determined by UPLC method with PDA detector at 310 nm， and then the apparent permeability coefficient Papp was calculated. The cytotoxic of different concentrations of ISO was investigated on Caco-2 cells to determine the concentration of drug administration. The effects of ISO concentration， time， temperature and transporter inhibitors on the transport of ISO were investigated. The test results showed that， ISO didn′t have significant cytotoxicity at 10-60 μmol·L-1 in 14 hours. The transportation of ISO on Caco-2 cells was related to the concentration to a certain extent. Papp of ISO was higher than 10×10-6 cm·s-1 and ISO was absorbed easily by Caco-2 cells. The transport volume of ISO at BL side reached maximum at 3 h and was slightly decreased at 6 h. Papp (AP-BL) and Papp(BL-AP) at 4 ℃ were lower than those at 37 ℃. Papp (AP-BL) of ISO was significantly increased after adding P-gp inhibitor verapamil and Papp (BL-AP) of ISO was significantly decreased after adding MRP-2 inhibitor (probenecid or MK-571). The results suggested that transport mode of ISO was mainly passive diffusion in Caco-2 cell models， and P-gp and MRP may be involved in the transport of ISO.
, 百拇医药
[Key words]Caco-2 cell monolayer model； isorhapontigenin； transport mechanism
異丹叶大黄素(isorhapontigenin，ISO)存在于买麻藤类植物中，具有良好的抗炎[1-2]，抗氧化[3-5]，治疗胰岛素抵抗及肥胖[6]等活性。近年来，有研究发现其有显著的抗膀胱癌活性[7-11]。有研究表明，小鼠灌胃不同浓度的异丹叶大黄素，采用LC-MS/MS测定其不同时间点的血药浓度，发现异丹叶大黄素在小鼠体内迅速达峰，消除迅速，代谢较快[9]。Caco-2细胞来源于人体结肠腺癌细胞，经培养分化后可形成与人体小肠上皮细胞一致的形态及功能，含有广泛存在于人体小肠上皮细胞的药物代谢酶及各种转运体[12]，可以用来研究药物在体外吸收和代谢情况。该模型是目前应用最广泛，最经典的用于研究药物吸收转运特性的体外模型。, 百拇医药(袁紫朔 张婷婷 金波 李彤 马辰)