Porcine milk-derived exosomes promote proliferation of intestinal epithelial cells.

Ting Chen,Yong-Liang Zhang,Rui-Ping Sun,Mei-Ying Xie,De-Lin Lin,Rui-Song Ye,Li-Min Wei,Jia-Jie Sun,Meng Li,Qing-Yan Jiang,Qian-Yun Xi,Xiao Cheng

doi:10.1038/srep33862

Ting Chen, Yong-Liang Zhang + Show 10 more

Open Access

https://doi.org/10.1038/srep33862

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Journal: Scientific Reports	Publication Date: Sep 20, 2016
Citations: 146	License type: CC BY 4.0

Affiliation: South China Agricultural University

Abstract

Milk-derived exosomes were identified as a novel mechanism of mother-to-child transmission of regulatory molecules, but their functions in intestinal tissues of neonates are not well-studied. Here, we characterized potential roles of porcine milk-derived exosomes in the intestinal tract. In vitro, treatment with milk-derived exosomes (27 ± 3 ng and 55 ± 5 ng total RNA) significantly promoted IPEC-J2 cell proliferation by MTT, CCK8, EdU fluorescence and EdU flow cytometry assays. The qRT-PCR and Western blot analyses indicated milk-derived exosomes (0.27 ± 0.03 μg total RNA) significantly promoted expression of CDX2, IGF-1R and PCNA, and inhibited p53 gene expression involved in intestinal proliferation. Additionally, six detected miRNAs were significantly increased in IPEC-J2 cell, while FAS and SERPINE were significantly down-regulated relative to that in control. In vivo, treated groups (0.125 μg and 0.25 μg total RNA) significantly raised mice’ villus height, crypt depth and ratio of villus length to crypt depth of intestinal tissues, significantly increased CDX2, PCNA and IGF-1R’ expression and significantly inhibited p53′ expression. Our study demonstrated that milk-derived exosomes can facilitate intestinal cell proliferation and intestinal tract development, thus giving a new insight for milk nutrition and newborn development and health.

Highlights

Were proposed to have the potential to influence the immune system of infants[14,15]
IPEC-J2 cells were seeded at a density between 4,000 to 12,000 cells per/well (96-well plate), and their growth peaked between 24 h to 72 h after seeding
Porcine milk-derived exosomes containing 27 ± 3 ng and 55 ± 5 ng total RNA were added to the IPEC-J2 cell culture, followed by analysis using the MTT assay at 0, 24, 48 and 72 h post treatment

Summary

Introduction

Were proposed to have the potential to influence the immune system of infants[14,15]. Even though milk-derived exosomes are known to be resistant to a harsh environment and are taken up by multiple cells, including intestinal cells, whether they regulate intestinal cell proliferation and intestinal tract development remains unclear. Based on our previous exploration of miRNAs, mRNAs and proteins in porcine milk[19], the KEGG pathway analysis of potential targets of top 10 miRNAs in porcine milk were enriched in Notch signaling pathway which could play an early critical role in cell fate determination and later roles in the regulation of cell proliferation and differentiation[20]. We hypothesized that porcine milk-derived exosomes are capable of regulating intestinal cell proliferation and intestinal tract development. We hope this study could provide a new evidence for the regulation of intestine tract development from breastmilk exosomes to offspring

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Conclusion