Extracellular Vesicle-like Associated microRNAs in Monofloral Honeys: Molecular Characterization and Functional...

Recent studies have identified microRNAs (miRNAs) in honey, opening a new and promising area of nutrition research. In this view, pasteurized and unpasteurized samples of Eucalyptus, Orange Blossom, Chestnut, and Sulla honeys were analyzed using manual and semi-automated RNA extraction methods. Semi-automated extraction yielded significantly higher RNA amounts than manual methods, while pasteurization selectively affected miRNA presence, depending on the type of honey. The panel of conserved miRNAs monitored was let-7a-5p, miR-1-3p, miR-7-5p, miR-10a-5p, miR-33a-5p, miR-34a-5p, miR-92a-3p, miR-125b-5p and miR-133a-3p, across honey varieties and in their extracellular vesicles with structures approximately 200 nm in diameter that retain four miRNAs in all honey types, miR-1-3p, miR-34a-5p, miR-92a-3p, and miR-133a-3p. Bioinformatic analyses of validated miRNA targets revealed enrichment in pathways related to cytoskeletal organization, transcriptional regulation, protein stability, and immune system processes, with Reactome categories clustering around signal transduction, protein metabolism, and immune interactions. Cell-type-specific enrichment suggested that gastric isthmus progenitor cells, stromal cells, and immune subsets could be potential targets, implying roles in epithelial renewal, immune modulation, and wound healing. Overall, these findings enhance our understanding of honey as a source of conserved miRNAs in extracellular vesicles, highlighting its potential as a natural carrier that protects miRNAs from degradation. This study offers new insights into the health-promoting properties of honey, warranting further preclinical studies.