Heat acclimation increases mitochondrial respiration capacity of C2C12 myotubes and protects against LPS-mediated energy deficit

  • Meghan G. Patton
  • Trevor L. Gillum
  • Mandy C. Szymanski
  • Lacey M. Gould
  • Claire J. Lauterbach
  • Roger A. Vaughan
  • Matthew R. Kuennen
Original Paper

Abstract

This work investigated the effect of a 6-day heat acclimation (HA) protocol on myotube metabolic responses at baseline and in response to a subsequent lipopolysaccharide (LPS) challenge. C2C12 myotubes were incubated for 2 h/day at 40 °C for 6 days (HA) or maintained at 37 °C (C). Following 24-h recovery, myotubes were challenged with 500 ng/ml LPS for 2 h, then collected for analysis of protein markers of mitochondrial biogenesis and macronutrient storage. Functional significance of these changes was confirmed with mitochondrial respiration and glycolytic measurements on a Seahorse XF-96 analyzer. HA stimulated mitochondrial biogenesis and increased indicators of mitochondrial content [SIRT1 (+ 62%); PGC-1α (+ 57%); NRF-1 (+ 40%); TFAM (+ 141%); CS (+ 25%); CytC (+ 38%); all p < 0.05]. Altered lipid biosynthesis enzymes [p-ACCa:ACC (+ 59%; p = 0.04) and FAS (− 86%; p < 0.01)] suggest fatty acid generation may have been downregulated, whereas increased GLUT4 (+ 69%; p < 0.01) and LDH-B (+ 366%; p < 0.01) suggest aerobic glycolytic capacity may have been improved. Mitochondrial biogenesis signaling in HA myotubes was suppressed by 500 ng/ml LPS (PGC-1α, NRF-1, TFAM; all p > 0.05) but increased LDH-B (+ 30%; p = 0.02) and CPT-1 (+ 55%; p < 0.01) suggesting improved catabolic function. Basal respiration was increased in HA myotubes (+ 8%; p < 0.01) and HA myotubes maintained elevated basal respiration during LPS challenge (+ 8%; p < 0.01). LPS reduced peak respiration in C myotubes (− 6%; p < 0.01) but did not impair peak respiration in HA myotubes (p > 0.05). Oxidative reliance was elevated in HA over that in control (+ 25%; p < 0.01) and in HA + LPS over C + LPS (+ 30%; p < 0.01). In summary, HA stimulated mitochondrial biogenesis in C2C12 myotubes. HA myotubes exhibited (1) elevated basal/peak mitochondrial respiration capacities; (2) greater oxidative reliance; and (3) protection against LPS-mediated respiration impairment. Collectively, these data suggest HA may improve aerobic metabolism in skeletal muscle and protect against LPS-mediated energy deficit.

Keywords

Hyperthermia Lipopolysaccharide Oxidative metabolism Oxidative reliance 

Abbreviations

p-ACC

Phosphorylated acetyl-CoA carboxylase

ACC

Acetyl-CoA carboxylase

p-AMPK

Phosphorylated 5′-AMP-activated protein kinase

AMPK

5′-AMP-activated protein kinase

CPT

Carnitine palmitoyltransferase I

CS

Citrate synthase

CytC

Cytochrome c

FAS

Fatty acid synthase

FOXO1

Forkhead in rhabdomyosarcoma

GLUT4

Glucose uptake transporter 4

GS

Glycogen synthase

p-GSK-3α

Phosphorylated glycogen synthase kinase 3 alpha

p-GSK-3β

Phosphorylated glycogen synthase kinase 3 beta

GSK-3α/β

Glycogen synthase kinase 3 alpha/beta

LDH-A

Lactate dehydrogenase A

LDH-B

Lactate dehydrogenase B

MYH4

Myosin heavy chain IIB

MyoD

Myocyte differentiation factor

TFAM

Mitochondrial transcription factor A

TNC

Troponin C

NRF-1

Nuclear respiratory factor 1

PGC-1α

Peroxisome proliferator-associated receptor gamma co-activator-1

PPAR β/δ

Peroxisome proliferator-associated receptor beta/delta

PPAR γ

Peroxisome proliferator-associated receptor gamma

SIRT1

Sirtuin 1

UCP2

Uncoupling protein 2

Notes

Acknowledgements

We would like to thank the Department of Physical Therapy (Congdon School of Health Sciences) and the School of Arts and Sciences for the use of shared lab space and equipment.

Author contribution

MGP made a significant contribution to the first draft of the manuscript and was lead author. MGP, MCS, LMG, and CJL assisted with experiments and manuscript preparation. TLG assisted with manuscript preparation. RAV designed and conducted experiments and assisted with manuscript preparation. MRK is corresponding author. He conceived the study, designed, conducted and oversaw all experiments, performed statistical analyses, oversaw MGP in drafting of the manuscript, and wrote subsequent versions of the manuscript as dictated by the reviewers. All authors read and approved the final manuscript.

Supplementary material

12192_2018_894_MOESM1_ESM.docx (278 kb)
Supplementary Fig. 1 (DOCX 277 kb)
12192_2018_894_MOESM2_ESM.docx (35 kb)
Supplemental Table 1 (DOCX 34 kb)

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Copyright information

© Cell Stress Society International 2018

Authors and Affiliations

  • Meghan G. Patton
    • 1
  • Trevor L. Gillum
    • 2
  • Mandy C. Szymanski
    • 1
  • Lacey M. Gould
    • 1
  • Claire J. Lauterbach
    • 1
  • Roger A. Vaughan
    • 1
  • Matthew R. Kuennen
    • 1
  1. 1.Department of Exercise ScienceHigh Point UniversityHigh PointUSA
  2. 2.Department of KinesiologyCalifornia Baptist UniversityRiversideUSA

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