Uptake Pathway of Apple-derived Nanoparticle by Intestinal Cells to Deliver its Cargo

Pharm Res. 2021 Mar;38(3):523-530. doi: 10.1007/s11095-021-03018-8. Epub 2021 Mar 15.


Purpose: Food-derived nanoparticles exert cytoprotective effects on intestinal cells by delivering their cargo, which includes macromolecules such as microRNAs and proteins, as well as low-molecular weight compounds. We previously reported that apple-derived nanoparticles (APNPs) downregulate the expression of human intestinal transporter OATP2B1/SLCO2B1 mRNA. To verify the involvement of the cargo of APNPs in affecting the expression of transporters, we characterized the uptake mechanism of APNPs in intestinal cells.

Methods: The uptake of fluorescent PKH26-labeled APNPs (PKH-APNPs) into Caco-2, LS180, and HT-29MTX cells was evaluated by confocal microscopy and flow cytometry.

Results: The uptake of PKH-APNPs was prevented in the presence of clathrin-dependent endocytosis inhibitors, chlorpromazine and Pitstop2. Furthermore, PKH-APNPs were incorporated by the HT29-MTX cells, despite the disturbance of the mucus layer. Additionally, the decrease in SLCO2B1 mRNA by APNPs was reversed by Pitstop 2 in Caco-2 cells, indicating that APNPs decrease SLCO2B1 by being incorporated via clathrin-dependent endocytosis.

Conclusions: We demonstrated that clathrin-dependent endocytosis was mainly involved in the uptake of APNPs by intestinal cells, and that the cargo in the APNPs downregulate the mRNA expression of SLCO2B1. Therefore, APNPs could be a useful tool to deliver large molecules such as microRNAs to intestinal cells.

Keywords: OATP2B1; apple-derived nanoparticles; clathrin-dependent endocytosis; food-drug interaction; intestinal uptake.

MeSH terms

  • Biological Transport
  • Caco-2 Cells
  • Clathrin / metabolism
  • Endocytosis
  • Fluorescent Dyes / chemistry
  • Gene Expression Regulation / drug effects
  • HT29 Cells
  • Humans
  • Intestines / cytology
  • Intestines / pathology*
  • Malus / chemistry*
  • Nanoparticles / chemistry*
  • Nanoparticles / metabolism*
  • Optical Imaging
  • Organic Anion Transporters / genetics
  • Organic Anion Transporters / metabolism
  • RNA, Messenger / metabolism
  • Signal Transduction
  • Tissue Distribution


  • Clathrin
  • Fluorescent Dyes
  • Organic Anion Transporters
  • RNA, Messenger
  • SLCO2B1 protein, human