黄芩苷及其固体脂质纳米粒在Caco-2细胞上的吸收机制研究

徐倩, 欧阳怡, 吴鸿飞

中国药学杂志 ›› 2019, Vol. 54 ›› Issue (12) : 1000-1006.

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中国药学杂志
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中国药学杂志 ›› 2019, Vol. 54 ›› Issue (12) : 1000-1006. DOI: 10.11669/cpj.2019.12.013
论著

黄芩苷及其固体脂质纳米粒在Caco-2细胞上的吸收机制研究

  • 徐倩, 欧阳怡, 吴鸿飞*
作者信息 +

Absorption Mechanism of Baicalin and Its Solid Lipid Nanoparticles on Caco-2 Cells

  • XU Qian, OUYANG Yi, WU Hong-fei*
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摘要

目的 通过建立体外Caco-2细胞模型研究黄芩苷(BA)及其固体脂质纳米粒(SLN)的吸收特性。方法 利用CCK-8法、LDH法筛选BA及BA SLN在Caco-2细胞单层模型的合适浓度。采用高效液相色谱法测定BA含量,分别研究时间、浓度、温度、内吞抑制剂及外排抑制剂对其摄取、转运量的影响;利用Western blot法检测外排蛋白的表达量。结果 在50～150 μg·mL-1,BA及BA SLN摄取呈浓度依赖性;在4～37 ℃,BA及BA SLN的细胞摄取量随温度升高而增加;内吞抑制剂影响BA SLN的细胞摄取量;多药耐药相关蛋白2(MRP2)抑制剂和乳腺癌耐药蛋白(BCRP)抑制剂显著降低BA外排率,对BA SLN外排率无影响;BA SLN降低细胞MRP2、BCRP表达量。结论 BA在Caco-2细胞模型的吸收为被动扩散,并伴随能量依赖,与MRP2、BCRP外排有关。BA SLN显著增加药物在细胞模型的吸收,可能与其增加内吞、抑制细胞外排蛋白表达减少药物外排有关。

Abstract

OBJECTIVE To study the absorption mechanism of baicalin (BA) and baicalin solid lipid nanoparticles (BA SLN) by establishing an in vitro Caco-2 cell model. METHODS A Caco-2 cell model was established. The appropriate concentration of BA and BA SLN in Caco-2 cell monolayer model was screened by CCK-8 method and LDH method. The content of BA was determined by high performance liquid chromatography, and the effects of time, concentration, temperature and endocytosis inhibitor on the uptake of BA and BA SLN were studied by this model. The transport of BA and BA SLN in the presence or absence of efflux inhibitors was also investigated. The expression of efflux protein was detected by Western blot. RESULTS At 50-150 μg·mL-1, the uptake of BA and BA SLN was concentration-dependent; at 4 to 37 ℃, the uptake of BA and BA SLN increased with increasing temperature; endocytosis inhibitor influenced the cellular uptake of BA SLN; multidrug resistance-associated protein 2 (MRP2) inhibitors and breast cancer resistance protein (BCRP) inhibitors significantly reduced BA efflux, but did not affect BA SLN efflux; blank SLN and BA SLN can reduce the expression of MRP2 and BCRP in cells. CONCLUSION BA is taken up and transported by small intestinal epithelial cells in a passive manner, and may be accompanied by energy dependence, which is related to MRP2 and BCRP efflux. BA SLN can significantly increase the uptake of drugs in the Caco-2 cell model, which may be related to the increase of endocytic pathway uptake and inhibition of extracellular repressor expression inhibition drug efflux.

关键词

黄芩苷 / 固体脂质纳米粒 / Caco-2细胞模型 / 摄取 / 转运 / 外排蛋白

Key words

baicalin / baicalin solid lipid nanoparticle / Caco-2 cell model / uptake / transport / efflux protein

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徐倩, 欧阳怡, 吴鸿飞. 黄芩苷及其固体脂质纳米粒在Caco-2细胞上的吸收机制研究[J]. 中国药学杂志, 2019, 54(12): 1000-1006 https://doi.org/10.11669/cpj.2019.12.013
XU Qian, OUYANG Yi, WU Hong-fei. Absorption Mechanism of Baicalin and Its Solid Lipid Nanoparticles on Caco-2 Cells[J]. Chinese Pharmaceutical Journal, 2019, 54(12): 1000-1006 https://doi.org/10.11669/cpj.2019.12.013
中图分类号： R944   

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基金

国家自然科学基金项目资助(81873038);安徽省教育厅高校优秀青年人才支持计划重点项目资助(gxyqZD2016135)

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