不同配伍比例的葛根素-甘草酸自组装纳米粒在Caco-2细胞单层模型的转运研究

Transport study of different proportions of puerarin-glycyrrhizic acid self-assembled nanoparticles in a Caco-2 cell monolayer model

关键词:  葛根素  甘草酸  自组装  纳米粒  Caco-2细胞模型  转运特性

Key words:  puerarin  glycyrrhizic acid  self-assembly  nanoparticle  Caco-2 cell model  transport characteristics

摘要:

[目的] 为提高葛根素的溶解度和肠道吸收能力，基于葛根-甘草药对煎煮过程中成分间相互作用，考察其主要成分葛根素（Pue）与甘草酸（GA）在不同摩尔比下制备的葛根素-甘草酸自组装纳米粒（Pue-GA NPs）在 Caco-2 细胞单层模型的吸收转运特性。[方法] 采用溶剂挥发法制备纳米粒，考察其粒径和形貌，通过紫外光谱和分子对接分析其相互作用。使用 Transwell 法比较物理混合物与自组装纳米粒及不同摩尔比纳米粒的跨膜转运差异，并结合4种内吞抑制剂探讨吸收机制。[结果] 不同摩尔比Pue-GA NPs均形成稳定的球形纳米粒，氢键作用参与其组装。Caco-2 细胞转运实验显示，摩尔比为1∶1的Pue-GA NPs跨膜转运能力最强，显著优于Pue-GA物理混合物，且4种内吞抑制剂均可降低其跨膜转运能力，以盐酸氯丙嗪（CPZ）最显著。[结论] GA与Pue可以高效自组装形成纳米粒。Pue-GA NPs显著提升Pue在Caco-2细胞的吸收转运效率。摩尔比为1∶1的 Pue-GA NPs 跨膜转运能力最强，吸收过程主要依赖网格蛋白介导的内吞途径。

Abstract:

[Objective] To enhance the solubility and intestinal absorption of puerarin（Pue），the study investigated the absorption and transport characteristics of self-assembled puerarin-glycyrrhizic acid nanoparticles（Pue-GA NPs） prepared at different molar ratios using a Caco-2 cell monolayer model. The research was inspired by the component interactions occurring during the decoction of the traditional Chinese herbal pair Radix Puerariae-Radix Glycyrrhizae. [Methods] Pue-GA NPs were prepared via a solvent evaporation method，and their particle size and morphology were characterized. The interactions between Pue and glycyrrhizic acid（GA） were evaluated through UV-visible spectroscopy and molecular docking. Transwell assays were employed to compare the transmembrane transport of physical mixtures versus self-assembled nanoparticles at different molar ratios. Additionally，four endocytic inhibitors were applied to elucidate the underlying absorption mechanisms. [Results] Pue-GA NPs with different molar ratios formed stable spherical nanoparticles，and hydrogen bonding was involved in their self-assembly. Caco-2 transport studies revealed that Pue-GA NPs at a 1∶1 molar ratio exhibited the highest transmembrane transport efficiency，which was significantly greater than that of the physical mixture. All four endocytic inhibitors reduced nanoparticle transport，with chlorpromazine showing the most pronounced inhibitory effect. [Conclusion] Glycyrrhizic acid and puerarin can efficiently self-assemble into stable nanoparticles. The Pue-GA NPs notably improved the transport efficiency of Pue across the Caco-2 cell monolayer，with the 1∶1 molar ratio formulation exhibiting the best performance. The enhanced absorption was primarily mediated via a clathrin-dependent endocytic pathway.