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Pharmacological HIF1 Inhibition Eliminates Downregulation of the Pentose Phosphate Pathway and Prevents Neuronal Apoptosis in Rat Hippocampus Caused by Severe Hypoxia

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Abstract

The pentose phosphate pathway (PPP) of glucose metabolism in the brain serves as a primary source of NADPH which in turn plays a crucial role in multiple cellular processes, including maintenance of redox homeostasis and antioxidant defense. In our model of protective mild hypobaric hypoxia in rats (3MHH), an inverse correlation between hypoxia-inducible factor-1 (HIF1) activity and mRNA levels of glucose-6-phosphate dehydrogenase (G6PD), the key enzyme of PPP, was observed. In the present study, it was demonstrated that severe hypobaric hypoxia (SH) induced short-term upregulation of HIF1 alpha-subunit (HIF1α) in the hippocampal CA1 subfield and decreased the activity of G6PD. The levels of NADPH were also reduced, promoting oxidative stress, triggering apoptosis, and neuronal loss. Injection of a HIF1 inhibitor (HIF1i), topotecan hydrochloride (5 mg/kg, i.p.), before SH prevented the upregulation of HIF1α and normalized G6PD activity. In addition, HIF1i injection caused an increase in NADPH levels, normalization of total glutathione levels and of the cellular redox status as well as suppression of free-radical and apoptotic processes. These results demonstrate a new molecular mechanism of post-hypoxic cerebral pathology development which involves HIF1-dependent PPP depletion and support a recently suggested injurious role of HIF1 activation in the acute phase of cerebral hypoxia/ischemia. Application of PPP stimulators in early post-hypoxic/ischemic period might represent a promising neuroprotective strategy.

HIF1-dependent down-regulation of the pentose phosphate pathway contributes to the hypoxia-induced oxidative stress and neuronal apoptosis in the rat hippocampus

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Abbreviations

G6PD:

Glucose-6-phosphate dehydrogenase

HIF1:

Hypoxia-inducible factor-1

MHH:

Mild hypobaric hypoxia

NADPH:

Reduced nicotinamide adenine dinucleotide phosphate

PPP:

Pentose phosphate pathway

SH:

Severe hypobaric hypoxia

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Acknowledgments

Instrumental analyses were performed at the Research park of Saint Petersburg State University, Research Centre for Environmental Safety and Resource Centre for Molecular and Cell Technologies. The work has been supported by Russian Foundation for Basic Research (RFBR) grant no. 19-015-00336 and Russian Program of State Academies GP-14 (section 65).

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Authors and Affiliations

  1. Laboratory of Regulation of Brain Neuron Functions, Pavlov Institute of Physiology, Russian Academy of Sciences, Makarova emb. 6, 199034, Saint Petersburg, Russia

    Oleg Vetrovoy, Kseniia Sarieva, Ekaterina Tyulkova & Elena Rybnikova

  2. Department of Biochemistry, Faculty of Biology, Saint Petersburg State University, Universitetskaya emb. 7-9, 199034, Saint Petersburg, Russia

    Oleg Vetrovoy, Natalia Eschenko, Olga Galkina & Andrey Lyanguzov

  3. Laboratory of Cell Biology in Culture, Institute of Cytology, Russian Academy of Sciences, Tihoretsky pr. 4, 194064, Saint Petersburg, Russia

    Ekaterina Lomert

  4. Institute for Regenerative Medicine, Medical Research and Education Center, Lomonosov Moscow State University, Lomonosov Ave. 27-10, 119192, Moscow, Russia

    Peter Nimiritsky

  5. Faculty of Medicine, Lomonosov Moscow State University, Lomonosov Ave. 31-5, 119192, Moscow, Russia

    Peter Nimiritsky

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  1. Oleg Vetrovoy
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  2. Kseniia Sarieva
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  8. Ekaterina Tyulkova
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  9. Elena Rybnikova
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Contributions

Oleg Vetrovoy was responsible for the experimental design, performed the experiments, and wrote the paper.

Kseniia Sarieva participated in the experiments, contributed to the experimental design and data analysis.

Ekaterina Tyulkova, Ekaterina Lomert, and Peter Nimiritsky contributed to the experiments.

Andrey Lyanguzov performed statistical analysis of the data.

Olga Galkina, Natalia Eschenko, and Elena Rybnikova contributed to the implementation of the study, data analysis, and preparation of the paper.

All authors read and approved the final manuscript.

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Correspondence to Oleg Vetrovoy.

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Vetrovoy, O., Sarieva, K., Lomert, E. et al. Pharmacological HIF1 Inhibition Eliminates Downregulation of the Pentose Phosphate Pathway and Prevents Neuronal Apoptosis in Rat Hippocampus Caused by Severe Hypoxia. J Mol Neurosci 70, 635–646 (2020). https://doi.org/10.1007/s12031-019-01469-8

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