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Assessing the survival of exogenous plant microRNA in mice.

Liang G, Zhu Y, Sun B, Shao Y, Jing A, Wang J, Xiao Z - Food Sci Nutr (2014)

Bottom Line: The survival of plant small RNAs from the diet in animals, however, remains unclear, and the persistence of miRNAs from dietary plants in the animal gastrointestinal (GI) tract is still under debate.Exogenous plant miRNAs were present in the sera, feces, and tissues of animals and these exogenous plant miRNAs were primarily acquired orally.The amount of miR-172 that survived passage through the GI tract varied among individuals, with a maximum of 4.5% recovered at the stomach of one individual, and had a range of 0.05-4.5% in different organs.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University Nanjing, 210096, China ; School of Medical Technology and Engineering, Henan University of Science and Technology Luo Yang 471003, Henan, China.

ABSTRACT
MicroRNAs (miRNAs), a class of small RNAs, are important molecules that influence several developmental processes and regulate RNA interference (RNAi), and are abundant in animals, plants, and plant tissues that are traditionally consumed in the diet. The survival of plant small RNAs from the diet in animals, however, remains unclear, and the persistence of miRNAs from dietary plants in the animal gastrointestinal (GI) tract is still under debate. In this study, ICR mice were fed plant total RNAs in quantities of 10-50 μg, extracted from Brassica oleracea. Serum, feces, and various tissues were collected from the mice after RNA consumption and analyzed for several miRNAs. Exogenous plant miRNAs were present in the sera, feces, and tissues of animals and these exogenous plant miRNAs were primarily acquired orally. MiR-172, the most highly enriched exogenous plant miRNA in B. oleracea, was found in the stomach, intestine, serum, and feces of mice that were fed plant RNA extracts including miR-172. The amount of miR-172 that survived passage through the GI tract varied among individuals, with a maximum of 4.5% recovered at the stomach of one individual, and had a range of 0.05-4.5% in different organs. Furthermore, miR-172 was detected in the blood, spleen, liver, and kidney of mice.

No MeSH data available.


Determination of mature miR-172 copy number. (A) Standard curves were constructed for mature miR-172 using synthetic miR-172 standard sample. Tenfold dilutions of miR-172 RNA oligos was quantified by real-time PCR. (B) The assays produced a 6-log dynamic range, lower limit of detection of 2 × 10 copies and an R2 > 0.99.
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fig04: Determination of mature miR-172 copy number. (A) Standard curves were constructed for mature miR-172 using synthetic miR-172 standard sample. Tenfold dilutions of miR-172 RNA oligos was quantified by real-time PCR. (B) The assays produced a 6-log dynamic range, lower limit of detection of 2 × 10 copies and an R2 > 0.99.

Mentions: Furthermore, we attempted to quantitate the proportion of the orally administered RNA that survived in the different samples with the standard curve (Fig. 4). The results of a TaqMan probe experiment suggested that of the order of 0.3–1.8% of the miR-172 originally administered could be recovered from the feces (Fig. 5C). By comparison to the results of reconstitution experiments it was determined that the stomach contained about 4.5–0.4% (2–24 h after feeding), the intestines 2.4–0.2% (2–36 h), blood about 1.3–0.2% (2–72 h), and spleen about 0.38–0.04% (2–72 h) of the miR-172 orally administered (Fig. 5). It is essential to note that, in some of the experiments, 30–50 μg of small RNA was added to the food provided to the mice. Obviously, under this feeding regime, the exact time of ingestion of small RNA could not be determined. We hence recorded the times of feces collection after offering the small RNA-treated food. In this case, miR-172 could be detected in the content of the stomach and intestines of mice at 2, 4, 9, 24, until 36 h. after presenting the food containing small RNA, but at 72 h after pellet exposure, the amount of miR-172 was significantly less.


Assessing the survival of exogenous plant microRNA in mice.

Liang G, Zhu Y, Sun B, Shao Y, Jing A, Wang J, Xiao Z - Food Sci Nutr (2014)

Determination of mature miR-172 copy number. (A) Standard curves were constructed for mature miR-172 using synthetic miR-172 standard sample. Tenfold dilutions of miR-172 RNA oligos was quantified by real-time PCR. (B) The assays produced a 6-log dynamic range, lower limit of detection of 2 × 10 copies and an R2 > 0.99.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221836&req=5

fig04: Determination of mature miR-172 copy number. (A) Standard curves were constructed for mature miR-172 using synthetic miR-172 standard sample. Tenfold dilutions of miR-172 RNA oligos was quantified by real-time PCR. (B) The assays produced a 6-log dynamic range, lower limit of detection of 2 × 10 copies and an R2 > 0.99.
Mentions: Furthermore, we attempted to quantitate the proportion of the orally administered RNA that survived in the different samples with the standard curve (Fig. 4). The results of a TaqMan probe experiment suggested that of the order of 0.3–1.8% of the miR-172 originally administered could be recovered from the feces (Fig. 5C). By comparison to the results of reconstitution experiments it was determined that the stomach contained about 4.5–0.4% (2–24 h after feeding), the intestines 2.4–0.2% (2–36 h), blood about 1.3–0.2% (2–72 h), and spleen about 0.38–0.04% (2–72 h) of the miR-172 orally administered (Fig. 5). It is essential to note that, in some of the experiments, 30–50 μg of small RNA was added to the food provided to the mice. Obviously, under this feeding regime, the exact time of ingestion of small RNA could not be determined. We hence recorded the times of feces collection after offering the small RNA-treated food. In this case, miR-172 could be detected in the content of the stomach and intestines of mice at 2, 4, 9, 24, until 36 h. after presenting the food containing small RNA, but at 72 h after pellet exposure, the amount of miR-172 was significantly less.

Bottom Line: The survival of plant small RNAs from the diet in animals, however, remains unclear, and the persistence of miRNAs from dietary plants in the animal gastrointestinal (GI) tract is still under debate.Exogenous plant miRNAs were present in the sera, feces, and tissues of animals and these exogenous plant miRNAs were primarily acquired orally.The amount of miR-172 that survived passage through the GI tract varied among individuals, with a maximum of 4.5% recovered at the stomach of one individual, and had a range of 0.05-4.5% in different organs.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University Nanjing, 210096, China ; School of Medical Technology and Engineering, Henan University of Science and Technology Luo Yang 471003, Henan, China.

ABSTRACT
MicroRNAs (miRNAs), a class of small RNAs, are important molecules that influence several developmental processes and regulate RNA interference (RNAi), and are abundant in animals, plants, and plant tissues that are traditionally consumed in the diet. The survival of plant small RNAs from the diet in animals, however, remains unclear, and the persistence of miRNAs from dietary plants in the animal gastrointestinal (GI) tract is still under debate. In this study, ICR mice were fed plant total RNAs in quantities of 10-50 μg, extracted from Brassica oleracea. Serum, feces, and various tissues were collected from the mice after RNA consumption and analyzed for several miRNAs. Exogenous plant miRNAs were present in the sera, feces, and tissues of animals and these exogenous plant miRNAs were primarily acquired orally. MiR-172, the most highly enriched exogenous plant miRNA in B. oleracea, was found in the stomach, intestine, serum, and feces of mice that were fed plant RNA extracts including miR-172. The amount of miR-172 that survived passage through the GI tract varied among individuals, with a maximum of 4.5% recovered at the stomach of one individual, and had a range of 0.05-4.5% in different organs. Furthermore, miR-172 was detected in the blood, spleen, liver, and kidney of mice.

No MeSH data available.