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HIF-P4H-2 deficiency protects against skeletal muscle ischemia-reperfusion injury

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Abstract

We show here that mice hypomorphic for hypoxia-inducible factor prolyl 4-hydroxylase-2 (HIF-P4H-2) (Hif-p4h-2 gt/gt), the main regulator of the stability of the HIFα subunits, have normoxic stabilization of HIF-1α and HIF-2α in their skeletal muscles. The size of the capillaries, but not their number, was increased in the skeletal muscles of the Hif-p4h-2 gt/gt mice, whereas the amount of glycogen was reduced. The expression levels of genes for glycolytic enzymes, glycogen branching enzyme 1 and monocarboxylate transporter 4, were increased in the Hif-p4h-2 gt/gt skeletal muscles, whereas no significant increases were detected in the levels of any vasculature-influencing factor studied. Serum lactate levels of the Hif-p4h-2 gt/gt mice recovered faster than those of the wild type following exercise. The Hif-p4h-2 gt/gt mice had elevated hepatic phosphoenolpyruvate carboxykinase activity, which may have contributed to the faster clearance of lactate. The Hif-p4h-2 gt/gt mice had smaller infarct size following limb ischemia-reperfusion injury. The increased capillary size correlated with the reduced infarct size. Following ischemia-reperfusion, glycogen content and ATP/ADP and CrP/Cr levels of the skeletal muscle of the Hif-p4h-2 gt/gt mice were higher than in the wild type. The higher glycogen content correlated with increased expression of phosphofructokinase messenger RNA (mRNA) and the increased ATP/ADP and CrP/Cr levels with reduced apoptosis, suggesting that HIF-P4H-2 deficiency supported energy metabolism during ischemia-reperfusion and protection against injury.

Key messages

  • HIF-P4H-2 deficiency protects skeletal muscle from ischemia-reperfusion injury.

  • The mechanisms involved are mediated via normoxic HIF-1α and HIF-2α stabilization.

  • HIF-P4H-2 deficiency increases capillary size but not number.

  • HIF-P4H-2 deficiency maintains energy metabolism during ischemia-reperfusion.

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Acknowledgments

We thank T. Aatsinki, R. Juntunen, A. Kokko, E. Lehtimäki, M. Siurua, the personnel of the Biocenter Oulu EM Core Facility co-funded by the University of Oulu and Biocenter Finland, and the University of Oulu Laboratory Animal Centre for their excellent technical assistance. This work was supported by Academy of Finland grants 120156, 140765, 218129, and 266719 to P.K., Academy of Finland grants 200471 and 202469 and Center of Excellence 2012–2017 grant 251314 to J.M.; grants from the S. Jusélius Foundation to P.K. and J.M.; a grant from the Emil Aaltonen Foundation to P.K.; grants from the Jane and Aatos Erkko Foundation to P.K. and J.M.; and a grant from FibroGen, Inc. to J.M.).

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Authors and Affiliations

  1. Biocenter Oulu, Faculty of Biochemistry and Molecular Medicine, Oulu Center for Cell-Matrix Research, University of Oulu, Oulu, Finland

    Sara Karsikas, Mikko Myllymäki, Minna Heikkilä, Kari I. Kivirikko, Johanna Myllyharju, Raisa Serpi & Peppi Koivunen

  2. Biocenter Oulu, Department of Pathology, University of Oulu, Oulu, Finland

    Raija Sormunen

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  1. Sara Karsikas
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Correspondence to Peppi Koivunen.

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Conflict of interest

K.I.K. is a scientific founder and consultant of FibroGen, Inc., which develops HIF-P4H inhibitors as potential therapeutics. K.I.K. and J.M. own equity in this company, and the company has sponsored research in the laboratory headed by K.I.K. and currently supports research headed by J.M. No other potential conflicts of interest relevant to this article were reported.

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Karsikas, S., Myllymäki, M., Heikkilä, M. et al. HIF-P4H-2 deficiency protects against skeletal muscle ischemia-reperfusion injury. J Mol Med 94, 301–310 (2016). https://doi.org/10.1007/s00109-015-1349-0

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  • DOI: https://doi.org/10.1007/s00109-015-1349-0

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