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Inflammatory markers CD11b, CD16, CD66b, CD68, myeloperoxidase and neutrophil elastase in eccentric exercised human skeletal muscles

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Abstract

The aim of the present study was to investigate leucocyte markers, CD11b, CD16, CD66b, CD68, myeloperoxidase and neutrophil elastase on skeletal muscle biopsies from biceps brachii after unaccustomed eccentric exercise followed by the second bout of exercise 3 weeks later. The subjects (10 subjects received COX-2 inhibitor (Celecoxib) and 13 subjects received placebo) were divided into three categories: mild, moderate and severe effect of eccentric exercise, according to the reduction and recovery of muscle force-generating capacity after performing 70 maximal eccentric actions with elbow flexors on an isokinetic dynamometer. The results showed that the CD66b antibody was applicable for localization of neutrophils in human skeletal muscle, whereas the other studied neutrophil markers recognized also other leucocytes than neutrophils. The number of CD66b positive cells in skeletal muscle was very low and was not affected by the exercise. The macrophage marker CD68 showed reactivity also against satellite cells and fibroblast-like cells in skeletal muscle and therefore cannot be applied as a quantitative value for inflammatory cells. Skeletal muscle fibre injury, shown as dystrophin negative fibres, was observed approximately in half of the biopsies at 4 and 7 days after the first exercise bout in the categories moderate and severe effect of eccentric exercise. These subjects represent the most prominent loss in muscle force-generating capacity both at the category and the individual levels. Furthermore, deformed skeletal muscle fibres were observed in five subjects in these categories after the second bout of exercise. The present results suggest that neutrophils are not involved in skeletal muscle fibre injury and the reduction in muscle force-generating capacity after a single bout of eccentric exercise is a good indirect indicator of muscle damage in humans. Furthermore, prolonged regeneration process could be one of the reasons for impaired peripheral muscle function after high-force eccentric exercise.

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Abbreviations

BSA:

Bovine serum albumin

CD68:

Cluster of differentiation 68

CEL S01:

Celecoxib subject 01

DAPI:

4′,6-Diamidino-2-phenylindole

g :

Gravity

M r :

Molecular weight

O.C.T:

Optimal cutting temperature

PBO S01:

Placebo subject 01

ROI:

Region of interest

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Acknowledgments

The authors would like to thank Prof. Lars Mørkrid (Department of Medical Biochemistry, University Hospital, Oslo, Norway) for the serum creatine kinase analyses. The authors will also thank Jan Gunnar Fjeld (Department of Nuclear Medicine at Oslo University Hospital, Oslo, Norway) for the scintigraphic measurements. This study was partly financed by Pfizer Inc. (Norway). However, the authors have no economical interests in Pfizer, nor are there any contract-bond restrictions or clauses that potentially could have influenced the study. The authors at the Department of Anatomy (University of Oslo, Norway) received financial support from the Norwegian Research Council.

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Authors and Affiliations

  1. Department of Physical Performance, Norwegian School of Sport Science, Sognsveien 220, 0863, Oslo, Norway

    Gøran Paulsen & Truls Raastad

  2. Department of Molecular Biosciences, University of Oslo, Blindernveien 31, 0371, Oslo, Norway

    Ingrid Egner

  3. Department of Pathology, University of Oslo and Oslo University Hospital, Sognsvannsveien 9, 0372, Oslo, Norway

    Finn Reinholt, Sverre-Henning Brorson & Satu Koskinen

  4. Department of Anatomy and Centre for Molecular Biology and Neuroscience, University of Oslo, Sognsvannsveien 9, 0372, Oslo, Norway

    Simen Owe & Fredrik Lauritzen

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  1. Gøran Paulsen
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  2. Ingrid Egner
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  3. Truls Raastad
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Correspondence to Satu Koskinen.

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Paulsen, G., Egner, I., Raastad, T. et al. Inflammatory markers CD11b, CD16, CD66b, CD68, myeloperoxidase and neutrophil elastase in eccentric exercised human skeletal muscles. Histochem Cell Biol 139, 691–715 (2013). https://doi.org/10.1007/s00418-012-1061-x

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