中国中药杂志

2019, v.44(01) 167-174

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白及有效部位中主要成分在Caco-2细胞中的吸收特性研究
Absorptive characteristic of Bletilla striata extract in Caco-2 cell monolayer

杨淑婷;潘洁;陆苑;梅朝叶;刘亭;黄勇;王永林;
YANG Shu-ting;PAN Jie;LU Yuan;MEI Chao-ye;LIU Ting;HUANG Yong;WANG Yong-lin;Guizhou Provincial Key Laboratory of Pharmaceutics,Guizhou Medcial University;School of Pharmacy,Guizhou Medical University;Engineering Research Center for the Development and Application of Ethnic Medicine and Traditional Chinese Medicine(Ministry of Education),Guizhou Medical University;State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University;

摘要(Abstract):

为研究白及有效部位中主要成分在Caco-2细胞中的吸收特性,初步阐明白及有效部位的口服吸收机制。该实验采用Caco-2细胞单层细胞吸收模型,通过UPLC-Q-TOF分析白及有效部位在Caco-2细胞中的摄取成分,并建立UPLC-TQD分析方法测定白及有效部位主要成分在Caco-2细胞中的含量,分别考察时间、浓度、pH和P-gp抑制剂对各成分的吸收影响。经过UPLC-Q-TOF指认白及有效部位中的主要成分B6,B12,B14,B17,B19,B23,研究发现该6个成分能够摄取进入Caco-2细胞中,并且在Caco-2细胞中的吸收呈现浓度依赖性。其中B12,B14的吸收在0~180 min为上升趋势,未呈现饱和,B6,B17,B19,B23在60 min后吸收趋于饱和;各成分在偏酸性或酸性条件下吸收较好,其中B6在pH 6.0环境中吸收较好,其余成分B12,B14,B17,B19,B23在pH 4.0环境中吸收较好;且加入维拉帕米P-gp抑制剂后和空白相比,B6,B12的吸收无差异,而B14,B17,B19,B23的Caco-2细胞摄取量增加,但无显著性差异,说明B6等6个成分的摄取过程中没有P-gp的参与,即B6等6个成分不是P-gp底物。由此结果可知白及有效部位主要成分进入Caco-2细胞以被动扩散为主,且吸收过程不受P-糖蛋白的外排作用影响。
To investigate the active fraction from Bletilla striata in Caco-2 cell monolayer,so as to explore its absorption mechanism of oral administration preliminarily.Active fraction from B.striata in Caco-2 cell monolayer was analyzed by UPLC-Q-TOF and detected by UPLC-MS/MS,and the effects of different concentrations,pH and P-glycoprotein inhibitors on Caco-2 cells Monolayer were investigated.Six compounds were isolated from the active fraction of B.striata in Caco-2 cell monolayer by UPLC-Q-TOF,and identified as B6,B12,B14,B17,B19 and B23,with concentration dependence.Within the 0-180 min,the uptake of B12 and B14 had a time dependence,while B6,B17,B19 and B23 tended to saturate after 60 min.All of the components had a good absorption in an acidic environment.B6 had a good absorption at pH 6.0,while the other components B12,B14,B17,B19 and B23 had a good absorption at pH4.0.The absorption of the 6 main components of B.striata were not be affected by P-glycoprotein inhibitors(verapamil/cyclosporin A).Compared with the control group,there was no difference in the absorption of B6 and B12,and the absorption of B14,B17,B19 and B23 increased,but with no significant difference.The absorption characteristic of B.striata extract across the Caco-2 cell monolayer is probably passive diffusion,and the absorption process was not affected by P-glycoprotein.

关键词(KeyWords): 白及;Caco-2细胞;UPLC-Q-TOF;UPLC-TQD;被动扩散;P-gp
Bletilla striata;Caco-2;UPLC-Q-TOF;UPLC-TQD;passive diffusion;P-glycoprotein

Abstract:

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基金项目(Foundation): 国家自然科学基金项目(81460630,81360636);; 贵州省科技计划项目(黔科合平台人才[2017]5601,[2016]5613/5677);; 贵阳市科研创新团队项目(筑科合同[2017]30-29)

作者(Author): 杨淑婷;潘洁;陆苑;梅朝叶;刘亭;黄勇;王永林;
YANG Shu-ting;PAN Jie;LU Yuan;MEI Chao-ye;LIU Ting;HUANG Yong;WANG Yong-lin;Guizhou Provincial Key Laboratory of Pharmaceutics,Guizhou Medcial University;School of Pharmacy,Guizhou Medical University;Engineering Research Center for the Development and Application of Ethnic Medicine and Traditional Chinese Medicine(Ministry of Education),Guizhou Medical University;State Key Laboratory of Functions and Applications of Medicinal Plants,Guizhou Medical University;

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