连翘苷经血脑屏障体外透过机制研究  

Study of Mechanism on in vitro Permeation of Phillyrin through Blood-brain Barrier

作　　者：黄海英[1,2] 苏成福[2] 郭艳丽[2] 魏永恒[1] 张北月 石晋丽[1] 

机构地区：[1]北京中医药大学中药学院,北京100029 [2]河南中医学院药学院,郑州450046

出　　处：《中国药房》2015年第22期3049-3051,共3页China Pharmacy

基　　金：国家重大新药创制(No.2012ZX09102201-18);河南中医学院科研苗圃工程(No.MP2013-15)

摘　　要：目的：研究连翘苷经血脑屏障的体外透过机制。方法：以0（阴性对照）、10、25、50、75、100μg/ml连翘苷溶液培养大肠癌MDR1基因转染的马丁达比犬肾上皮（MDCK-MDR1）细胞24 h,刃天青法检测细胞活力并计算细胞生存率。以10、25、50、75、100μg/ml连翘苷溶液培养MDCK-MDR1细胞10 min后,检测胞内连翘苷含量并绘制质量浓度-摄取速率曲线;0（阴性对照）、50、100μg/ml连翘苷溶液培养MDCK-MDR1细胞3 h后,倒置荧光显微镜下观察细胞紧密连接蛋白结构。结果：与阴性对照比较,10~100μg/ml连翘苷溶液培养细胞24 h后细胞存活率无明显改变。在10~100μg/ml质量浓度范围内,MDCK-MDR1细胞摄取速率与连翘苷的质量浓度不呈线性关系,有逐渐饱和趋势。50、100μg/ml连翘苷溶液培养细胞3 h后,细胞紧密联接蛋白完整。结论：在透过模拟血脑屏障时,连翘苷的吸收可能为被动转运结合主动转运,其质量浓度对细胞紧密联接蛋白有影响。OBJECTIVE：To research the mechanism of in vitro permeation of phillyrin through blood-brain barrier.METHODS：After Madin-Darby canine kidney epithelial cells transfected with colorectal cancer MDR1 gene（MDCK-MDR1）were cultured with phillyrin solution of 0（negative control）,10,25,50,75 and 100 μg/ml for 24 h,cell viability was determined by resazurin method and cell survival rate was calculated.After MDCK-MDR1 cells were cultured with phillyrin solution of 10,25,50,75 and 100 μg/ml for 10 min,the content of phillyrin in the cells was determined,and concentration-uptake rate curve was drawn.Following 3 h culture of MDCK-MDR1 cells with phillyrin solution of 0（negative control）,50 and 100 μ g/ml,the structure of cell tight junction protein was observed under the inverted microscope.RESULTS：Compared to the negative control group,after24 h cell culture with phillyrin solution of 10-100 μg/ml,no obvious change in cell survival rate occurred.MDCK-MDR1 cells cultured with the phillyrin at a mass concentration of 10-100 μ g/ml demonstrated a nonlinear relationship with concentration of phillyrin and a gradual saturation trend.After the cells were cultured with phillyrin of 50 and 100 μg/ml for 3 h,cell tight junction protein was intact.CONCLUSIONS：The absorption of phillyrin through the simulated blood-brain barrier may be in the form of passive transportation combined with active transportation,the concertration has effect on cell tight junction protein.

关 键 词：MDR1基因转染 马丁达比犬肾上皮细胞 血脑屏障 连翘苷 细胞紧密联接蛋白 体外试验 

分 类 号：R965[医药卫生—药理学] R285[医药卫生—药学]