岩白菜素在MDCK-MDR1细胞模型中的跨膜转运机制  

Transport mechanism of bergenin aross MDCK-MDR1 cell monolayer model

作　　者：吴秀稳 温然 王娜[1] 李丽 王苑宇[2] WU Xiu-wen;WEN Ran;WANG Na;LI Li;WANG Yuan-yu(School of Pharmaceutical Sciences,Hebei Medical University,Shijiazhuang 050017,China;The Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学药学院,河北石家庄050017 [2]河北医科大学第二医院,河北石家庄050000

出　　处：《中国药理学通报》2022年第11期1698-1704,共7页Chinese Pharmacological Bulletin

基　　金：河北省自然科学基金资助项目(No.H2019206249,H2019206205);河北省高等学校科学技术研究项目(No.QN2019121,QN2019118);河北省医学科学研究课题计划(No.20200934)。

摘　　要：目的研究岩白菜素透血脑屏障(blood-brain barrier,BBB)的转运机制。方法采用MTT法考察岩白菜素的无毒剂量范围;采用分子对接法预测岩白菜素与P-糖蛋白(P-glycoprotein,P-gp)的相互作用(结合方式及作用力);采用体外MDCK-MDR1细胞模型模拟岩白菜素的透BBB跨膜转运,并分析透过-浓度关系、透过-时间关系以及P-gp抑制剂维拉帕米对其跨膜转运的影响。结果在5~40μmol·L^(-1)浓度范围内岩白菜素对MDCK-MDR1细胞无毒性;岩白菜素主要通过氢键和疏水作用与P-gp结合,复合物P-gp-岩白菜素的稳定性强于P-gp-维拉帕米;岩白菜素从顶(apical,AP)端到底(basolateral,BL)端转运的表观渗透系数(P_(app AP→BL))为(1.07±0.27)×10^(-6) cm·s^(-1)(t=90 min,c=20μmol·L^(-1)),外排率(efflux rate,ER)为4.32;岩白菜素与维拉帕米联合给药,岩白菜素的P_(app AP→BL)明显增大(P<0.01),P_(app BL→AP)明显减小(P<0.05),ER值减小至0.98。结论岩白菜素为透BBB中等的化合物,其跨膜转运过程除被动扩散外,还有P-gp的参与。Aim To study the transport mechanism of bergenin passing through blood-brain barrier(BBB).Methods MTT assay was used to investigate the toxicity of bergenin on MDCK-MDR1 cells.Molecular docking was used to predict the binding mode and effect ability of bergenin with P-gp.In vitro MDCK-MDR1 cell monolayer model was used to analyze transport characteristics of bergenin and the effect of concentration,time and verapamil(a P-gp inhibitor)on the transport of bergenin.Results Bergenin was non-toxic to MDCK-MDR1 cells within the concentration of 5 to 40μmol·L^(-1).There was hydrogen-bond and hydrophobic interaction between P-gp and bergenin,and P-gp-bergenin was more stable than P-gp-verapamil.The P_(app) value of bergenin transported from AP to BL(P_(app AP→BL))was(1.07±0.27)×10^(-6) cm·s^(-1) at 20μmol·L^(-1) for a period of 90 min;addition of verapamil,P_(app AP→BL) values of bergenin increased significantly(P<0.01),while those of transport in the opposite direction decreased significantly(P<0.05);correspondingly,the efflux ratio(ER)for bergenin was decreased from 4.32 to 0.98.Conclusions Bergenin has moderate BBB permeability,and its transport process is driven by not only passive diffusion,but also P-gp.

关 键 词：岩白菜素 透血脑屏障 MDCK-MDR1细胞模型 P-糖蛋白 分子对接 转运机制 

分 类 号：R284.1[医药卫生—中药学] R322.61[医药卫生—中医学] R322.81R341.32