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Measuring liver triglyceride content in mice: non-invasive magnetic resonance methods as an alternative to histopathology

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Abstract

Object

Quantitative assessment of liver fat is highly relevant to preclinical liver research and should ideally be performed non-invasively. This study aimed to compare three non-invasive Magnetic Resonance (MR) and two histopathological methods against the reference standard of biochemically determined liver triglyceride content (LTC).

Materials and methods

A total of 50 mice [21 C57Bl/6OlaHsd mice (C57Bl/6), nine low-density lipoprotein (LDL) receptor knock-out −/− (LDL −/−) mice and 20 C57BL/6 mice] received either a high-fat, high-fat-high-cholesterol or control diet, respectively. Mice were examined 4, 8 or 12 weeks into the diet using MR [1H-MR Spectroscopy, Proton Density Fat Fraction (PDFF), mDixon] and histopathological methods (visual scoring or digital image analysis (DIA) of Oil-Red-O (ORO) stained liver sections). Correlations [Pearson’s coefficient (r)] were studied with respect to LTC.

Results

Microvesicular steatosis was seen in 42/50 mice. 1H-MRS values showed normal to moderately elevated liver fat content. Visual scoring and DIA of ORO-sections correlated moderately with LTC at r = 0.59 and r = 0.49 (P < 0.001), respectively. 1H-MRS, PDFF and mDixon correlated significantly better, at r = 0.74, r = 0.75 and r = 0.82, respectively.

Conclusion

Non-invasively determined MR measures of normal to moderately elevated liver fat in mice had a higher correlation with LTC than invasive histopathological measures. Where available, MR is the preferred method for fat quantification.

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Abbreviations

DIA:

Digital image analysis

H&E:

Haematoxylin and eosin

LTC:

Liver triglyceride content

MRI:

Magnetic Resonance Imaging

1H-MRS:

1H Magnetic Resonance Spectroscopy

NAFLD:

Non-alcoholic fatty liver disease

NAS:

NAFLD Activity Score

NASH:

Non-alcoholic steatohepatitis

ORO:

Oil Red-O

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Acknowledgments

The authors thank N.M.A. Tiel and ARIA-personnel for assistance with animal handling, G.C.M. Grinwis MD, Ph.D. for discussions on mice pathology, M. Froeling Ph.D. for assistance with the DIA–ORO, C.B. Sirlin MD for distributing the Lipoquant plugin for OsiriX, and K.M. Moerman Ph.D. for assistance with revising the manuscript.

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Authors and Affiliations

  1. Department of Radiology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, 1105AZ, Amsterdam, The Netherlands

    Jurgen H. Runge, Aart J. Nederveen & Jaap Stoker

  2. Department of Pathology, Academic Medical Center, University of Amsterdam, Meibergdreef 9, Amsterdam, The Netherlands

    Pieter J. Bakker & Joanne Verheij

  3. Tytgat Institute for Liver and Intestinal Research, Academic Medical Center, University of Amsterdam, Meibergdreef 71, Amsterdam, The Netherlands

    Ingrid C. Gaemers & Theo B. M. Hakvoort

  4. Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Meibergdreef 9, Amsterdam, The Netherlands

    Roelof Ottenhoff

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  1. Jurgen H. Runge
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Correspondence to Jurgen H. Runge.

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Runge, J.H., Bakker, P.J., Gaemers, I.C. et al. Measuring liver triglyceride content in mice: non-invasive magnetic resonance methods as an alternative to histopathology. Magn Reson Mater Phy 27, 317–327 (2014). https://doi.org/10.1007/s10334-013-0414-3

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  • DOI: https://doi.org/10.1007/s10334-013-0414-3

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