摘要
目的研究P-糖蛋白(P-gp)对槲皮苷肠道吸收过程中跨膜转运和代谢转化的影响。方法采用Caco-2细胞摄取模型和单层细胞转运模型,按LC-MS法测定槲皮苷、槲皮素、异鼠李亭和柽柳黄素,研究P-gp对槲皮苷在肠道吸收过程中的作用及作用机制。结果胞内摄取:在不同孵育浓度槲皮苷组(3.0、9.0和27.0 mg/L)和P-gp抑制剂环孢素A(Cys A)共孵育组,胞内槲皮苷均呈现150 min内随时间先升后降,60 min达峰,120 min达较低水平稳态的趋势特征,两组之间有显著差异(P<0.05);各组均检测到槲皮素,但仅27.0 mg/L浓度组胞内槲皮素呈现与其原形糖苷相似的趋势特征,其余各组均未呈现浓度和时间依赖性;同时槲皮苷27.0 mg/L时,槲皮苷组和共孵育组均检到异鼠李亭和柽柳黄素,表现出与原型糖苷相似的趋势特征,但差异无显著性(P>0.05),3.0和9.0 mg/L浓度组未测到。跨膜转运:在跨膜转运过程中,槲皮苷组和Cys A组,150 min内基底侧槲皮苷含量均呈现持续上升的趋势特征,两组之间无显著差异;槲皮素、异鼠李亭和柽柳黄素均未达检测限。结论槲皮苷原型可被Caco-2细胞吸收并透过Caco-2细胞单层,在Caco-2细胞内和Caco-2单层细胞基底侧均可发生脱糖基作用和进一步的代谢转化,导致跨膜转运和胞内摄取动态特征的不同;P-gp对槲皮苷有外排作用,但对其代谢转化无影响。
Objective To explore the role of P-glycoprotein（P-gp）on cellular uptake and metabolism in the transmembrane transport of quercitrin. Methods Caco-2 cell monolayer and P-gp inhibitor Cyclosporin A（Cys A）were used in the study. Quercitrin,quercetin,isorhamnetin and tamarixetin were determined by LC-MS to study cellular uptake and metabolism of quercitrin on Caco-2 cells. Results Uptake of quercitrin by Caco-2 cells：in different concentration groups of quercitrin coincubating with and without Cys A,intracellular accumulation presented the following characteristics：the amount of quercitrin first rose,reached the peak in 60 min and then declined to a steady-state in 120 min. And meanwhile there were significant differences between the two different processing groups incubating with and without Cys A（P0.05）;quercetin was detected in all groups（3.0,9.0 and 27.0 mg/L）. But in the higher concentration groups incubating with and without Cys A,the intracellular quercetin presented a characteristics similar to its original glycosides and showed a significant difference（P0.05）,while the other groups showed no concentration-and time-dependence. At the same time,isorhamnetin and tamarixetin were detected in two higher concentration groups incubating with and without Cys A,which showed the trend similar to the original glycosides but no significant difference was obtained between the two processing groups（P0.05）. Isorhamnetin and tamarixetin were not detected in the low and middle concentration groups. Transmembrane transport：on the basal lateral of all groups,the content of the quercitrin in 150 min incubation time showed a trend of continuous rise,and there was no significant difference between the two processing groups. Quercetin,isorhamnetin and tamarixetin were not detected. Conclusion Intact quercitrin could be absorbed into the Caco-2 cells and transported across the Caco-2 cell monolayer,and suffered a series of further metabolism in the Caco-2 cells and the basal side of Caco-2 cell monolayer,leading to different characteristics between intracellular accumulation and transmembrane transport. P-gp reduces the transmembrane transport of quercitrin by its efflux function,but did not involved in quercitrin metabolism.
作者
李素云
李峥
高蔚娜
李敬来
郭长江
张振清
LI Su-yun1,2,LI Zheng3,GAO Wei-na2,LI Jing-lai3,GUO Chang-jiang2,ZHANG Zhen-qing3(1. Department of Clinical Nutrition, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, China; 2. Department of Nutrition, lnsitute of Environmental and Operational Medicine, Academy of Military Medical Sciences, Academy of Milltary Sciences, Tianjing 300050, China ; 3. Department of Drug metabolism, Insitute of Pharmacology and Toxicology, Academy of Military Medical Sciences, Academy of Military Sciences, Beijing 100850, China)
出处
《国际药学研究杂志》
CSCD
北大核心
2017年第12期1150-1154,1162,共6页
Journal of International Pharmaceutical Research
基金
国家自然科学基金面上项目(81372988)
