Analysis of Serum microRNA Expression Profiles and Comparison with Small Intestinal microRNA Expression Profiles in Weaned Piglets

Weaning stress induces tissue injuries and impairs health and growth in piglets, especially during the first week post-weaning. MicroRNAs (miRNAs) play vital roles in regulating stresses and diseases. Our previous study found multiple differentially expressed miRNAs in small intestine of piglets at four days post-weaning. To better understand the roles of miRNAs during weaning stress, we analyzed the serum miRNA expressional profile in weaned piglets (at four days post-weaning) and in suckling piglets (control) of the same age using miRNA microarray technology. We detected a total of 300 expressed miRNAs, 179 miRNAs of which were differentially expressed between the two groups. The miRNA microarray results were validated by RT-qPCR. The biological functions of these differentially expressed miRNAs were predicted by GO terms and KEGG pathway annotations. We identified 10 highly expressed miRNAs in weaned piglets including miR-31, miR-205, and miR-21 (upregulated) and miR-144, miR-30c-5p, miR-363, miR-194a, miR-186, miR-150, and miR-194b-5p (downregulated). Additionally, miR-194b-5p expression was significantly downregulated in serum and small intestine of weaned piglets. Our results suggest that weaning stress affects serum miRNA profiles in piglets. And serum miR-194b-5p levels can reflect its expressional changes in small intestine of piglets by weaning stress.

[1]  X. Tao,et al.  Transient effects of weaning on the health of newly weaning piglets , 2016 .

[2]  Kimihiro Hino,et al.  Inducible expression of microRNA-194 is regulated by HNF-1alpha during intestinal epithelial cell differentiation. , 2008, RNA.

[3]  Meredith B. Baker,et al.  MiR-21 is induced in endothelial cells by shear stress and modulates apoptosis and eNOS activity. , 2010, Biochemical and biophysical research communications.

[4]  F. Middleton,et al.  Alterations in serum microRNA in humans with alcohol use disorders impact cell proliferation and cell death pathways and predict structural and functional changes in brain , 2015, BMC Neuroscience.

[5]  X. Tao,et al.  MicroRNA Transcriptome in Swine Small Intestine during Weaning Stress , 2013, PloS one.

[6]  T. Luedde,et al.  Circulating MicroRNA-150 Serum Levels Predict Survival in Patients with Critical Illness and Sepsis , 2013, PloS one.

[7]  Yankai Xia,et al.  Specific serum microRNA profile in the molecular diagnosis of Hirschsprung's disease , 2014, Journal of cellular and molecular medicine.

[8]  B. Burwinkel,et al.  Cancer diagnosis and prognosis decoded by blood-based circulating microRNA signatures , 2013, Front. Genet..

[9]  J. Mendell,et al.  MicroRNAs in Stress Signaling and Human Disease , 2012, Cell.

[10]  A. Tijsen,et al.  Coronary Artery Disease CAD is caused by atherosclerosis of the coronary arteries , 2012 .

[11]  Yongqing Hou,et al.  Fish oil alleviates activation of the hypothalamic-pituitary-adrenal axis associated with inhibition of TLR4 and NOD signaling pathways in weaned piglets after a lipopolysaccharide challenge. , 2013, The Journal of nutrition.

[12]  Takashi Nakamura,et al.  Plasma and synovial fluid microRNAs as potential biomarkers of rheumatoid arthritis and osteoarthritis , 2010, Arthritis research & therapy.

[13]  Daniel B. Martin,et al.  Circulating microRNAs as stable blood-based markers for cancer detection , 2008, Proceedings of the National Academy of Sciences.

[14]  Shuhong Zhao,et al.  Discovery of Porcine microRNAs and Profiling from Skeletal Muscle Tissues during Development , 2008, PloS one.

[15]  Xi Chen,et al.  Serum microRNA expression profile as a biomarker in the diagnosis and prognosis of pancreatic cancer. , 2012, Clinical chemistry.

[16]  Jian-ning Zhang,et al.  miR-21 improves the neurological outcome after traumatic brain injury in rats , 2014, Scientific Reports.

[17]  Terence P. Speed,et al.  A comparison of normalization methods for high density oligonucleotide array data based on variance and bias , 2003, Bioinform..

[18]  Xiao-rong Zhan,et al.  MicroRNA-185 regulates expression of lipid metabolism genes and improves insulin sensitivity in mice with non-alcoholic fatty liver disease. , 2014, World journal of gastroenterology.

[19]  Maarten Hulsmans,et al.  MicroRNA-containing microvesicles regulating inflammation in association with atherosclerotic disease. , 2013, Cardiovascular research.

[20]  J. Yin,et al.  Birth oxidative stress and the development of an antioxidant system in newborn piglets , 2013, Free radical research.

[21]  Michael T. McManus,et al.  Identification of MiR-205 As a MicroRNA That Is Highly Expressed in Medullary Thymic Epithelial Cells , 2015, PloS one.

[22]  T. Blondal,et al.  Assessing sample and miRNA profile quality in serum and plasma or other biofluids. , 2013, Methods.

[23]  H. Weiner,et al.  MicroRNA-21, T helper lineage and autoimmunity , 2015, Oncotarget.

[24]  K. Livak,et al.  Real-time quantification of microRNAs by stem–loop RT–PCR , 2005, Nucleic acids research.

[25]  M. Lindsay,et al.  microRNAs and the immune response. , 2008, Trends in immunology.

[26]  N. Rajewsky,et al.  Deciphering the porcine intestinal microRNA transcriptome , 2010, BMC Genomics.

[27]  P. Dash,et al.  Human traumatic brain injury alters plasma microRNA levels. , 2010, Journal of neurotrauma.

[28]  Catalin Vasilescu,et al.  MicroRNA Fingerprints Identify miR-150 as a Plasma Prognostic Marker in Patients with Sepsis , 2009, PloS one.

[29]  Onnop Srivannavit,et al.  microParaflo biochip for nucleic acid and protein analysis. , 2007, Methods in molecular biology.

[30]  B. Li,et al.  Clinical evaluation of circulating microRNA-25 level change in sepsis and its potential relationship with oxidative stress. , 2015, International journal of clinical and experimental pathology.

[31]  J. Sundquist,et al.  Circulating microRNA-144-5p is associated with depressive disorders , 2015, Clinical Epigenetics.

[32]  X. Chen,et al.  Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases , 2008, Cell Research.

[33]  J. Nickels,et al.  Identification of miR-185 as a regulator of de novo cholesterol biosynthesis and low density lipoprotein uptake , 2014, Journal of Lipid Research.

[34]  R. Regazzi,et al.  Circulating microRNAs as novel biomarkers for diabetes mellitus , 2013, Nature Reviews Endocrinology.

[35]  Shuang Huang,et al.  Serum levels of microRNAs can specifically predict liver injury of chronic hepatitis B. , 2012, World journal of gastroenterology.

[36]  Qi Zheng,et al.  GOEAST: a web-based software toolkit for Gene Ontology enrichment analysis , 2008, Nucleic Acids Res..

[37]  S. Papageorgiou,et al.  The role of microRNAs in normal and malignant hematopoiesis , 2010, European journal of haematology.

[38]  M-L Si,et al.  MicroRNA-185 Targets SOCS3 to Inhibit Beta-Cell Dysfunction in Diabetes , 2015, PloS one.

[39]  M. Simionescu,et al.  MiR-486 and miR-92a Identified in Circulating HDL Discriminate between Stable and Vulnerable Coronary Artery Disease Patients , 2015, PloS one.

[40]  Yariv Yogev,et al.  Serum MicroRNAs Are Promising Novel Biomarkers , 2008, PloS one.

[41]  S. Sugiharto,et al.  Plasma metabolomic profiles and immune responses of piglets after weaning and challenge with E. coli , 2014, Journal of Animal Science and Biotechnology.

[42]  Claudio Franceschi,et al.  Age-related differences in the expression of circulating microRNAs: miR-21 as a new circulating marker of inflammaging , 2012, Mechanisms of Ageing and Development.

[43]  M. Wolzt,et al.  In vivo profile of the human leukocyte microRNA response to endotoxemia. , 2009, Biochemical and biophysical research communications.

[44]  J. Crenshaw,et al.  The biological stress of early weaned piglets , 2013, Journal of Animal Science and Biotechnology.

[45]  Growth factor TGF-β induces intestinal epithelial cell (IEC-6) differentiation: miR-146b as a regulatory component in the negative feedback loop , 2012, Genes & Nutrition.

[46]  Y. Naito,et al.  Muscle-enriched microRNA miR-486 decreases in circulation in response to exercise in young men , 2013, Front. Physiol..

[47]  J. Pluske.  Feed- and feed additives-related aspects of gut health and development in weanling pigs , 2013, Journal of Animal Science and Biotechnology.

[48]  Yongwei Yu,et al.  MicroRNA-410-5p as a potential serum biomarker for the diagnosis of prostate cancer , 2016, Cancer Cell International.

[49]  L. Pfeffer,et al.  The Type I IFN-Induced miRNA, miR-21 , 2015, Pharmaceuticals.

[50]  P. Brennan,et al.  Circulating MicroRNAs as Non-Invasive Biomarkers for Early Detection of Non-Small-Cell Lung Cancer , 2015, PloS one.

[51]  Chunxiang Zhang,et al.  MicroRNA-21 in Cardiovascular Disease , 2010, Journal of cardiovascular translational research.

[52]  N. Pottier,et al.  Increased Circulating miR-21 Levels Are Associated with Kidney Fibrosis , 2013, PloS one.

[53]  Yulong Yin,et al.  Hydrogen peroxide-induced oxidative stress activates NF-κB and Nrf2/Keap1 signals and triggers autophagy in piglets , 2015 .

[54]  P. Stothard,et al.  Potential Regulatory Role of MicroRNAs in the Development of Bovine Gastrointestinal Tract during Early Life , 2014, PloS one.