Identification of differentially expressed miRNAs after Lactobacillus reuteri treatment in the ileum mucosa of piglets

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Genes & Genomics (2020) 42:1327–1338 https://doi.org/10.1007/s13258-020-00998-6

RESEARCH ARTICLE

Identification of differentially expressed miRNAs after Lactobacillus reuteri treatment in the ileum mucosa of piglets Qi Wang1,2,3   · Qian Sun1 · Jing Wang1,2,3 · Xiaoyu Qiu1 · Renli Qi1,2,3 · Jinxiu Huang1,2,3 Received: 21 July 2020 / Accepted: 10 September 2020 / Published online: 26 September 2020 © The Genetics Society of Korea 2020

Abstract Background  Lactobacillus reuteri I5007 possesses many excellent probiotic characteristics in piglets. miRNA plays important role in host-microbiota interactions, but the mechanism by which L. reuteri I5007 regulates intestinal function through its influence on miRNA expression is unknown. Objective  This study analyzed the miRNA expression patterns in the ileum mucosa tissue of piglets by L. reuteri I5007 treatment, aim to clarify its molecular mechanism for regulating intestinal function through miRNA. Methods  Neonatal piglets were orally administered L. reuteri I5007 or a placebo daily starting on day 1, and differential expression of ileal miRNAs was analyzed at 10 and 20 days of age by small RNA sequencing. Results  361 known porcine miRNAs were identified, and ten miRNAs were highly expressed in the ileum mucosa in both treatments. Nineteen differentially expressed (DE) miRNAs were identified in response to L. reuteri treatment, and four DE miRNAs (ssc-miR-196a, -196b-5p, -1285 and -10386) were differentially expressed at both time points. The KEGG pathway analyses showed the targets of 19 DE miRNAs were involved in 63 significantly enriched pathways, including the PI3KAkt and MAPK pathways, which were confirmed to play important roles in probiotic-host communication. L. reuteri I5007 exerted anti-inflammatory effects by influencing the levels of inflammatory cytokines. Suppressor of cytokine signalling 4 gene was the target gene of ssc-miR-196a/-196b-5p, overexpression of ssc-miR-196a/-196b-5p downregulated the mRNA expression of IL-1β and TNFα in IPEC-J2 cells. Conclusion  Our study provides new insight into the role of miRNAs in the intestinal function of piglets after L. reuteri I5007 treatment. Keywords  Lactobacillus reuteri · miRNA · Differentially expressed · Ileum mucosa · Piglet

Introduction

Electronic supplementary material  The online version of this article (https​://doi.org/10.1007/s1325​8-020-00998​-6) contains supplementary material, which is available to authorized users. * Jinxiu Huang [email protected] 1



Chongqing Academy of Animal Sciences, Rongchang, Chongqing 402460, China

2



Key Laboratory of Pig Industry Sciences, Ministry of Agriculture, Rongchang, Chongqing 402460, China

3

Chongqing Key Laboratory of Pig Industry Sciences, Rongchang, Chongqing 402460, China



microRNAs (miRNAs) are a class of noncoding RNA molecules with lengths of 18– 25 nt, which take part in the regulation of mRNA expression at the post-transcriptional level (Fabian et al. 2010). During the last 20 years, plenty of studies have indicated that miRNAs participat