Abstract
The current study seeks to resolve the discrepancy in the literature regarding the cross-kingdom transfer of plant microRNAs (miRNAs) into mammals using an improved miRNA processing and detection method. Two studies utilizing C57BL/6 mice were performed. In the first study, mice were fed an AIN-93M diet and gavaged with water, random deoxynucleotide triphosphates (dNTP) or isolated corn miRNAs for two weeks (n = 10 per group). In the second study, mice were fed an AIN-93M diet, or the diet supplemented with 3% fresh or autoclaved corn powder for two weeks (n = 10 per group). Corn miRNA levels were analyzed in blood and tissue samples by real-time PCR (RT-PCR) following periodate oxidation and β elimination treatments to eliminate artifacts. After removing false positive detections, there were no differences in corn miRNA levels between control and treated groups in cecal, fecal, liver and blood samples. Using an in vitro digestion system, corn miRNAs in AIN-93M diet or in the extracts were found to be extensively degraded. Less than 1% was recovered in the gastrointestinal tract after oral and gastric phases. In conclusion, no evidence of increased levels of corn miRNAs in whole blood or tissues after supplementation of corn miRNAs in the diet was observed in a mouse model.
Highlights
MicroRNA are a group of small, functional, non-protein coding RNA oligonucleotides that were discovered two decades ago and are universally found in microorganisms, plants, and animals [1,2]
Taking advantage of 30 protective property of plant miRNA, we previously reported that periodate oxidation and β elimination reaction treatment targets the unmethylated 30 end of miRNAs to eliminate the artifacts in PCR detection of plant miRNAs in biological samples [20]
No significant reduction of corn miRNA in the diet was detected between the start and end of the study (Supplementary Materials, Figure S1)
Summary
MicroRNA (miRNA) are a group of small, functional, non-protein coding RNA oligonucleotides that were discovered two decades ago and are universally found in microorganisms, plants, and animals [1,2]. In 2012, Zhang and colleagues reported a novel cross-kingdom uptake of intact rice miRNAs (miR156a and miR168a) via dietary consumption into circulation and organs of humans and mice [6]. LDLRAP1 in the liver [6] This result suggests ingestion of plant miRNA may influence physiology and health in humans. The importance of this finding was signified by the number and breadth of follow-up studies [7,8,9,10,11,12,13,14,15,16,17,18]. No definitive conclusion has been reached regarding the extent and prevalence of dietary plant miRNAs entering circulation
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