Plant miRNA bol-miR159 Regulates Gut Microbiota Composition in Mice: In Vivo Evidence of the Crosstalk between Plant miRNAs and Intestinal Microbes

Abstract

New evidence reveals that bol-miR159, an miRNA rich in fruits and vegetables, cross-kingdomly functions in mammalian bodies. However, whether the miRNA could regulate gut microbiota remains unclear. Here, the effect of miR159 on mouse intestinal microbes was comprehensively examined. The results showed that supplementation of miR159 to the chow diet significantly enhanced the diversity of mouse gut microbiota without causing pathological lesions or inflammatory responses on the intestines. At the phylum level, miR159 increased the abundance of Proteobacteria and decreased the Firmicute-to-Bacteroidetes (F/ B) ratio. miR159 had prebiotic-like effects on mouse gut microbiota, as it promoted the growth of the bacteria that is beneficial for maintaining gut health. The miRNA can target bacteria genes and get into the bacteria cells. The data provide direct in vivo evidence on the crosstalk between plant miRNAs and intestinal microbes, highlighting the potential for miRNA-based strategies that modulate gut microbes to improve host health.

Atypical Plant miRNA cal-miR2911: Robust Stability against Food Digestion and Specific Promoting Effect on Bifidobacterium in Mice

“…Previous studies found that miR159 could survive in vitro digestion and significantly increase the diversity and richness of mouse gut microbiota. 12,17 Given its high were first subjected to acclimatization of 3 days and subjected to gavage feeding of saline (CD group), miR2911 oligomer (miR2911 group), nucleotides of miR2911 (NT group), and heteroduplex of miR2911 (HDmiR group). The mice were given the gavage once a day for 7 days, anesthetized, and sacrificed on the eighth day.…”

“…Previous studies found that miR159 could survive in vitro digestion and significantly increase the diversity and richness of mouse gut microbiota. , Given its high stability, we asked if miR2911 has a function of shaping the profile of intestinal microbes. The experiment scenario is shown in Figure A.…”

“…The sequencing was performed according to a previous report with slight modifications . Briefly, bacterial DNA was extracted using a Mag-Bind Soil DNA Kit (Omega Bio-Tek, Doraville, USA), and the purity was measured on a NanoDrop 2000 UV–vis spectrophotometer (Thermo Fisher Scientific, Wilmington, USA).…”

“…The assay was preformed according to a previous study with slight modifications . Briefly, miR2911-targeting sequences (wild and mutant) flanked by restriction sites were synthesized beforehand.…”

“…The sequencing was performed according to a previous report with slight modifications. 17 Briefly, bacterial DNA was extracted using a Mag-Bind Soil DNA Kit (Omega Bio-Tek, Doraville, USA), and the purity was measured on a NanoDrop 2000 UV−vis spectrophotometer (Thermo Fisher Scientific, Wilmington, USA). To generate DNA sequences carrying genetic information of the gut microbes, PCR amplification on the V3−V4 region of the 16S rRNA gene was performed using a forward primer (338F, 5′-ACTCCTACGGGAGG-CAGCAG-3′) and reverse primer (806R, 5′-GGAC-TACHVGGGTWTCTAAT-3′) containing degenerate bases.…”