Uptake of MicroRNAs from Exosome-Like Nanovesicles of Edible Plant Juice by Rat Enterocytes.

Yuko Ito,Yuki Kuranaga,Kazuhisa Uchiyama,Kohei Taniguchi,Sang-Woong Lee,Yosuke Inomata,Nabil Eid

doi:10.3390/ijms22073749

Abstract

MicroRNAs (miRNAs) are small RNAs present in extracellular vesicles (EVs) that, when transferred to a target cell, affect its biological functions. Plant miRNAs regulate the expression of certain mammalian genes. Here, we characterized EVs in fruit and vegetable juice, and their miRNA cargo, and investigated whether such miRNA-containing EVs could be taken up by mammalian enterocytes in vitro. Using filtration and ultra-centrifugation methods, EVs were purified from commercially available and manually squeezed plant juice. EV morphological features and subcellular localization were analyzed using the NanoSight tracking system and electron microscopy. Plant EV miRNA levels were evaluated using quantitative reverse transcription PCR. For the in vitro EV uptake experiments, rat intestinal epithelial cells (IEC6) were used. Plant EVs shared morphological features with mammalian EVs and contained miR156a-5p, miR166a-3p, and miR168a-5p. EVs were present in the cell sap-filled central vacuoles and were taken up by IEC6 cells. Edible plant cells produce EVs that contain various miRNAs and release them into the central vacuole. The exogenous plant EVs are taken up by mammalian enterocytes in vitro. These findings suggest the possibility that exogenous plant miRNAs carried by EVs can be absorbed via the gastrointestinal tract.

Highlights

MicroRNAs are small non-coding RNA molecules that are, on average, 25 nucleotides long
To characterize extracellular vesicles (EVs) in plant juice, NTA was first used to measure the size of plantTo characterize EVs in plant juice, NTA was first used to measure the size of plantderived EVs in paprika juice, pineapple juice
Smaller vesicles were observed in grapefruit SJ (Figure 2h). These analyses indicate that epidermal cells of grapefruit juice vesicles contain EVs

Summary

Introduction

MicroRNAs (miRNAs) are small non-coding RNA molecules that are, on average, 25 nucleotides long. They post-transcriptionally control the expression of the target gene by binding to the 30 -untranslated region of mRNA. MiRNAs from plant and animal food are absorbed by mammals [1]. As recently discovered, such food-derived miRNAs are involved in cross-kingdom gene regulation [2]. Plant miR168a targets the expression of the mammalian low-density lipoprotein receptor adapter protein 1 [3]. Mammalian cells secrete extracellular vesicles (EVs), which are intercellular communication tools and contain functional nucleic acids (mRNA, miRNA, or other RNA species)

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