人参皂苷靶向P糖蛋白增强紫杉醇对结肠癌细胞抑制作用  

Ginsenosides targeting P-glycoprotein enhance the inhibitory effect of paclitaxel on colon cancer

作　　者：朱晓慧 赵媛媛 李南熹 郭金南 田云飞 翟慧婷 王珊珊 杨德宣 窦桂芳 冯素香[1] 孟志云 ZHU Xiaohui;ZHAO Yuanyuan;LI Nanxi;GUO Jinnan;TIAN Yunfei;ZHAI Huiting;WANG Shanshan;YANG Dexuan;DOU Guifang;FENG Suxiang;MENG Zhiyun(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450000,China;Academy of Military Medical Sciences,Beijing 100850,China)

机构地区：[1]河南中医药大学药学院,河南郑州450000 [2]军事医学研究院,北京100850

出　　处：《中国药理学与毒理学杂志》2025年第2期89-99,共11页Chinese Journal of Pharmacology and Toxicology

基　　金：中药品种创新关键技术研究(145AHQ080014008X1202)。

摘　　要：目的探讨人参皂苷类P糖蛋白(P-gp)底物与紫杉醇联用对结肠癌Caco-2细胞增殖和迁移能力的影响。方法采用生物层干涉技术检测人参皂苷与P-gp的亲和力常数;采用网络分子对接预测人参皂苷与P-gp的亲合力;Caco-2细胞分为紫杉醇(0,6.25,12.5,25,50,100和200 mg·L^(-1))组、人参皂苷Rg3(0,6.25,12.5,25,50,100和200 mg·L^(-1))组及紫杉醇(5 mg·L^(-1))+人参皂苷Rg3(0,25,50,100和200 mg·L^(-1))组,分别与相应浓度化合物于37℃孵育48 h后,MTT法检测Caco-2细胞增殖抑制率,用“一带一线”法定量评价两药联用的相互作用;Caco-2细胞分为细胞对照组、紫杉醇5 mg·L^(-1)组、人参皂苷Rg350和100 mg·L^(-1)组及紫杉醇5 mg·L^(-1)+人参皂苷Rg350和100 mg·L^(-1)组,分别与相应浓度化合物于37℃孵育24 h后,分别采用集落实验及Transwell迁移实验检测细胞增殖和迁移能力;Caco-2细胞分为细胞对照组、奎尼丁12.5 mg·L^(-1)组及人参皂苷Rg36.25和12.5 mg·L^(-1)组,分别与相应浓度化合物于37℃孵育4 h后,采用ELISA检测P-gp及总蛋白表达量。结果人参皂苷Rb1,Rg3和Rg5与P-gp的亲和力常数均小于10-3mol·L^(-1),人参皂苷CK与P-gp的亲和力常数是10-2mol·L^(-1),人参皂苷Re与P-gp没有典型的结合解离曲线;人参皂苷Rg3,Rg5与P-gp的亲合力绝对值分别为8.5和7.6 kcal·mol-1;人参皂苷Rg3与紫杉醇(5 mg·L^(-1))联用通过协同及相加的方式抑制结肠癌细胞增殖,其中协同效应的剂量范围是[0+5,43.15+5]mg·L^(-1)和[164.51+5,200+5]mg·L^(-1),相加效应剂量的范围是[43.15+5,164.51+5]mg·L^(-1);与紫杉醇单用相比,两药联用后Caco-2细胞的增殖与迁移能力显著降低(均P<0.01);与细胞对照组相比,人参皂苷Rg36.25和12.5 mg·L^(-1)组及奎尼丁12.5 mg·L^(-1)组细胞总蛋白和P-gp表达量均显著降低(P<0.05)。结论人参皂苷通过与P-gp结合并抑制其活性,与紫杉醇协同降低Caco-2细胞的增殖和迁移能力。人参皂苷与紫杉醇�OBJECTIVE To investigate the effects of ginsenosides as P-glycoprotein(P-gp)substrates in combination with paclitaxel on the proliferation and migration of colon cancer Caco-2 cells.METHODS Bio-layer interferometry(BLI)technology was used to detect the constants of ginsenosides and P-gp.Network molecular docking was adopted to predict the binding affinity energy of ginsenosides and P-gp.Caco-2 cells were divided into paclitaxel 0,6.25,12.5,25,50,100 and 200 mg·L^(-1)groups,ginsenoside Rg30,6.25,12.5,25,50,100 and 200 mg·L^(-1)groups,and paclitaxel 5 mg·L^(-1)+ginsenoside Rg30,25,50,100 and 200 mg·L^(-1)groups.After 48 h of incubation,the growth inhibition rate of Caco-2 cells was detected by MTT assay,and the interaction between the two drugs was quantitatively evaluated using the"one-belt,one-line"modle.Caco-2 cells were divided into the cell control group,paclitaxel 5 mg·L^(-1)group,ginsenoside Rg350 and 100 mg·L^(-1)groups,and paclitaxel 5 mg·L^(-1)+ginsenoside Rg350 and 100 mg·L^(-1)groups.After 24 h of incubation,the proliferation and migration ability of the cells were detected by colony assay and Transwell migration assay.Caco-2 cells were then divided into the cell control group,quinidine 12.5 mg·L^(-1)group,and ginsenoside Rg36.25 and 12.5 mg·L^(-1)groups.After 4 h of incubation,the expression levels of P-gp and total protein were detected by ELISA.RESULTS The affinity constants of ginsenoside Rb1,Rg3,Rg5 with P-gp were all less than 10-3 mol·L^(-1),while that of ginsenoside CK with P-gp was 10-2 mol·L^(-1).There was no typical binding dissociation curve between ginsenoside Re and P-gp.The absolute binding affinities of ginsenosides Rg3 and Rg5 to P-gp were determined to be 8.5 kcal·mol-1 and 7.6 kcal·mol-1,respectively.Ginsenosides mixed with PTX 5 mg·L^(-1)inhibited the growth of colon cancer cells through synergy and addition,and the dose range of the synergistic effect was[0+5,43.15+5]mg·L^(-1);[164.51+5,200+5]mg·L^(-1),the additive effect dose ranged from[43.15+5,164.51+5]mg·L^(-1)

关 键 词：紫杉醇 人参皂苷 P糖蛋白 联合用药 

分 类 号：R966[医药卫生—药理学]