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A sustained increase in plasma NEFA upregulates the Toll-like receptor network in human muscle

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Abstract

Aims/hypothesis

Insulin-sensitive tissues (muscle, liver) of individuals with obesity and type 2 diabetes mellitus are in a state of low-grade inflammation, characterised by increased Toll-like receptor (TLR) expression and TLR-driven signalling. However, the cause of this mild inflammatory state is unclear. We tested the hypothesis that a prolonged mild increase in plasma NEFA will increase TLR expression and TLR-driven signalling (nuclear factor κB [NFκB] and mitogen-activated kinase [MAPK]) and impair insulin action in muscle of lean healthy individuals.

Methods

Twelve lean, normal-glucose-tolerant participants were randomised to receive a 48 h infusion (30 ml/h) of saline or Intralipid followed by a euglycaemic–hyperinsulinaemic clamp. Vastus lateralis muscle biopsies were performed before and during the clamp.

Results

Lipid infusion impaired insulin-stimulated IRS-1 tyrosine phosphorylation and reduced peripheral insulin sensitivity (p < 0.01). The elevation in circulating NEFA increased expression of TLR3, TLR4 and TLR5, and several MAPK (MAPK8, MAP4K4, MAP2K3) and inhibitor of κB kinase-NFκB (CHUK [IKKA], c-REL [REL] and p65 [RELA, NFKB3, p65]) signalling genes (p < 0.05). The lipid infusion also increased extracellular signal-regulated kinase (ERK) phosphorylation (p < 0.05) and tended to reduce the content of inhibitor of kappa Bα (p = 0.09). The muscle content of most diacylglycerol, ceramide and acylcarnitine species was unaffected. In summary, insulin resistance induced by prolonged low-dose lipid infusion occurs together with increased TLR-driven inflammatory signalling and impaired insulin-stimulated IRS-1 tyrosine phosphorylation.

Conclusions/interpretation

A sustained, mild elevation in plasma NEFA is sufficient to increase TLR expression and TLR-driven signalling (NFκB and MAPK) in lean individuals. The activation of this pathway by NEFA may be involved in the pathogenesis of insulin resistance in humans.

Trial registration ClinicalTrials.gov NCT01740817

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Abbreviations

DAG:

Diacylglycerol

ERK:

Extracellular signal-regulated kinase

ESI-MS/MS:

Electrospray tandem mass spectrometry

GSK-3:

Glycogen synthase kinase-3

HOMA-IR:

HOMA of insulin resistance

IκBα:

Inhibitor of kappa Bα

IKK:

Inhibitor of κB kinase

JNK:

c-Jun N-terminal kinase

LC-MS/MS:

High-performance liquid chromatography/mass spectrometry

M :

Insulin-stimulated glucose metabolism

MAPK:

Mitogen-activated kinase

NFκB:

Nuclear factor κB

PI3K:

Phosphatidylinositol 3-kinase

TLR:

Toll-like receptor

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Acknowledgements

We thank all the study participants and their families for their time and participation.

Funding

This work was supported by grants from the National Institutes of Health (NIH) (RO1-DK80157 and RO1-DK089229) and the American Diabetes Association to NM. This grant was supported by a University of Texas Health Science Center at San Antonio Clinical and Translational Science Award (TR000149) and an NIH National Service Research Award, Parent F32 (1F32DK095565-01A1) to SH. The research was supported in part by the Lipidomics Shared Resource, Hollings Cancer Center, Medical University of South Carolina (MUSC) (P30 CA138313) and the Lipidomics Core in the South Carolina Lipidomics and Pathobiology COBRE, Department Biochemistry, MUSC (P20 RR017677). This project also was supported by the National Center for Research Resources and the Office of the Director of the NIH (C06 RR018823) to MUSC.

Duality of interest

The authors declare that there is no duality of interest associated with this manuscript.

Contribution statement

SEH, HL, AA, YC, JGG, LA, JD and NM provided substantial contributions to the conception and design of this study and to the acquisition of data or analysis and interpretation of data. All authors were involved in drafting the article or revising it critically for important intellectual content and gave final approval of the version to be published.

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Authors and Affiliations

  1. Department of Medicine-Diabetes Division, University of Texas Health Science Center at San Antonio, San Antonio, TX, 78229, USA

    Sophie E. Hussey, Helen Lum, Andrea Alvarez, Yolanda Cipriani, Jesús Garduño-Garcia, Luis Anaya & Nicolas Musi

  2. Department of Medicine, The University of Pittsburgh, Pittsburgh, PA, USA

    John Dube

  3. Center for Healthy Aging, The University of Texas Health Science Center at San Antonio, San Antonio, TX, USA

    Nicolas Musi

  4. The Geriatric Research Education and Clinical Center, South Texas Veterans Health Care System, San Antonio, TX, USA

    Nicolas Musi

Authors
  1. Sophie E. Hussey
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  2. Helen Lum
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  3. Andrea Alvarez
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  4. Yolanda Cipriani
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  5. Jesús Garduño-Garcia
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  6. Luis Anaya
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  7. John Dube
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  8. Nicolas Musi
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Corresponding author

Correspondence to Nicolas Musi.

Additional information

Sophie E. Hussey and Helen Lum contributed equally to this work.

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Hussey, S.E., Lum, H., Alvarez, A. et al. A sustained increase in plasma NEFA upregulates the Toll-like receptor network in human muscle. Diabetologia 57, 582–591 (2014). https://doi.org/10.1007/s00125-013-3111-x

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