Ma Yi, Chen Xiaojiao, Diao Tiwei, Leng Yinjiang, Lai Xiaoqin, Wei Xin
College of Biological Engineering, Sichuan University of Science and Engineering, 188 University Town, Yibin 644000, China.
Foods. 2022 Oct 21;11(20):3299. doi: 10.3390/foods11203299.
This work aims to evaluate the effect of ferulic acid-grafted chitosan (FA-g-CS) on the interaction between anthocyanin (ANC) and / and their functions in ANC transmembrane transport using Caco-2 cells. The transmembrane transport experiments of ANC showed its low transport efficiency (Papp < 10 cm/s), whereas the phenomenon of a significantly rise in anthocyanins transport efficiency was observed with the incubation of FA-g-CS ( < 0.05). In order to investigate the mechanism of FA-g-CS improving ANC transmembrane transport, Caco-2 cells were transfected with small interfering RNA () specific for transporters and , and incubated with ANC, FA-g-CS, or their combination. Subsequently, Western blot analyses and immunofluorescence staining were carried out to monitor the intracellular and levels. These siRNA-transfected cells, incubated with compounds, indicate that and participated in the ANC transmembrane transport and that FA-g-CS, ANC, or their combination enhance / expression. In particular, Caco-2 cells incubated with both FA-g-CS and ANC show significantly increased or expression (>80%) compared with exclusively using FA-g-CS or ANC (<60%). Molecular docking results demonstrate that there is a good binding between FA-g-CS/ANC and or . These results highlight that FA-g-CS promotes the transmembrane transport of ANC by influencing the interaction between ANC and /; the interaction between FA-g-CS and ANC could be another key factor that improves the bioavailability of ANC.
本研究旨在利用Caco-2细胞评估阿魏酸接枝壳聚糖(FA-g-CS)对花青素(ANC)之间相互作用及其在ANC跨膜转运中功能的影响。ANC的跨膜转运实验表明其转运效率较低(表观渗透系数Papp<10 cm/s),而在FA-g-CS孵育时观察到花青素转运效率显著提高的现象(P<0.05)。为了研究FA-g-CS改善ANC跨膜转运的机制,用针对转运蛋白 和 的小干扰RNA(siRNA)转染Caco-2细胞,并与ANC、FA-g-CS或它们的组合一起孵育。随后,进行蛋白质免疫印迹分析和免疫荧光染色以监测细胞内 和 的水平。这些用化合物孵育的siRNA转染细胞表明, 和 参与了ANC的跨膜转运,并且FA-g-CS、ANC或它们的组合增强了 / 的表达。特别是,与单独使用FA-g-CS或ANC相比(<60%),同时用FA-g-CS和ANC孵育的Caco-2细胞显示 或 表达显著增加(>80%)。分子对接结果表明FA-g-CS/ANC与 或 之间存在良好的结合。这些结果突出表明,FA-g-CS通过影响ANC与 / 之间的相互作用促进ANC的跨膜转运;FA-g-CS与ANC之间的相互作用可能是提高ANC生物利用度的另一个关键因素。