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Hepatic expression of inflammatory genes and microRNAs in pigs with high “cholesteryl ester transfer protein” (CETP) activity

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Abstract

Human obesity and obesity-related diseases (ORD) are growing health problems worldwide and represent a major public health challenge. Most of these diseases are complex conditions, influenced by many genes (including microRNAs) and environmental factors. Many metabolic perturbations are associated with obesity; e.g., low levels of high-density lipoproteins (HDL) are high risk factors of cardiovascular events. A number of genetic, lifestyle, and environmental factors have been shown to contribute to the lowering of HDL-cholesterol. One of these factors is cholesteryl ester transfer protein (CETP) promoting the redistribution of cholesteryl esters, triglycerides, and phospholipids between plasma proteins. Moreover, obesity and ORD are often linked with chronic low-grade inflammation leading to insulin resistance and endothelial and microvascular dysfunctions. The aim of this study was to detect differences in the hepatic expression of genes involved in low-grade inflammation and of obesity- and cholesterol-related microRNAs in two mixed breed populations of pigs (Yorkshire-Göttingen minipig, YM and Duroc-Göttingen minipig, DM) including males and females, with extreme phenotypes for CETP activity levels (designated as CETP-high and CETP-low, respectively). Furthermore, breed and gender differences were also investigated. We found significant difference (P < 0.05) in hepatic expression levels of several mRNAs and microRNAs between the CETP-high and -low groups (C5, IL1RN, IL18, and miR-223-5p); between the two mixed breeds (IL1RAP and miR-140-5p); and between gender (APOA1, IL1RN, and FBLN1). Furthermore, when taking breed into account we show that the transcriptional levels of TNF, miR20a, miR33b, and miR130a differed between the two CETP groups. We conclude that increased CETP activity is accompanied by a modest differential hepatic expression of several microRNAs and inflammatory-related genes. Furthermore, our study demonstrates that when modeling the analysis of expression data, it is important to take gender- and breed-specific effects into account.

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Acknowledgments

We would like to thank laboratory technician Karin Tarp for excellent assistance in the Fluidigm platform. We would like to thank Caroline M. Mentzel for statistical advice. We would also like to thank Dr. Philippe Lesnik for the CETP measurements in the F2 pig population.

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Authors and Affiliations

  1. Department of Veterinary Clinical and Animal Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark

    Susanna Cirera, Benedicte C. Juul Tørsleff & Merete Fredholm

  2. Section of Immunology and Vaccinology, Innate Immunology Group, National Veterinary Institute, Technical University of Denmark, Bülowsvej 27, 1870, Frederiksberg C, Denmark

    Benedicte C. Juul Tørsleff, Peter M. H. Heegaard & Kerstin Skovgaard

  3. Department of Nutrition, Exercise and Sports, Faculty of Science, University of Copenhagen, Copenhagen, Denmark

    Christian Ritz

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  1. Susanna Cirera
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  2. Benedicte C. Juul Tørsleff
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  5. Peter M. H. Heegaard
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Correspondence to Susanna Cirera.

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Supplementary Table 1

mRNA assays selected for this study. Ref, gene used as reference gene; F, forward primer; R, reverse primer. Ampliqon size, size of the product in the qPCR. Previously validated primer assays, targeting transcripts coding for several phase proteins and cytokines involved in inflammation, were selected for the present study. Supplementary material 1 (XLSX 15 kb)

Supplementary Table 2

microRNA assays selected for this study. F, forward primer; R, reverse primer. Reference, article used for the selection of microRNAs to investigate. Previously validated primer assays, targeting microRNAs involved in cholesterol metabolism, innate immune response, inflammation and obesity were selected for the present study. Supplementary material 2 (XLSX 15 kb)

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Cirera, S., Tørsleff, B.C.J., Ritz, C. et al. Hepatic expression of inflammatory genes and microRNAs in pigs with high “cholesteryl ester transfer protein” (CETP) activity. Mamm Genome 27, 503–510 (2016). https://doi.org/10.1007/s00335-016-9649-4

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