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Hypothermia Attenuates Neurotoxic Microglial Activation via TRPV4

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Abstract

Therapeutic hypothermia (TH) provides neuroprotection. However, the cellular mechanisms underlying the neuroprotective effects of TH are not fully elucidated. Regulation of microglial activation has the potential to treat a variety of nervous system diseases. Transient receptor potential vanilloid 4 (TRPV4), a nonselective cation channel, is activated by temperature stimulus at 27–35 °C. Although it is speculated that TRPV4 is associated with the neuroprotective mechanisms of TH, the role of TRPV4 in the neuroprotective effects of TH is not well understood. In the present study, we investigated whether hypothermia attenuates microglial activation via TRPV4 channels. Cultured microglia were incubated under normothermic (37 °C) or hypothermic (33.5 °C) conditions following lipopolysaccharide (LPS) stimulation. Hypothermic conditions suppressed the expression of pro-inflammatory cytokines, inducible nitric oxide synthase, and the number of phagocytic microglia. AMP-activated protein kinase (AMPK)–NF-κB signaling was inhibited under hypothermic conditions. Furthermore, hypothermia reduced neuronal damage induced by LPS-treated microglial cells. Treatment with TRPV4 antagonist in normothermic culture replicated the suppressive effects of hypothermia on microglial activation and microglia-induced neuronal damage. In contrast, treatment with a TRPV4 agonist in hypothermic culture reversed the suppressive effect of hypothermia. These findings suggest that TH suppresses microglial activation and microglia-induced neuronal damage via the TRPV4-AMPK–NF-κB pathway. Although more validation is needed to consider differences according to age, sex, and specific central nervous system regions, our findings may offer a novel therapeutic approach to complement TH.

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Abbreviations

AMPK:

AMP-activated protein kinase

DMEM:

Dulbecco’s modified Eagle’s medium

ERK:

Extracellular signal–regulated kinase

iNOS:

Inducible nitric oxide synthase

LPS:

Lipopolysaccharide

MAP-2:

Microtubule-associated protein-2

MCM:

Microglia conditioned medium

RT-PCR:

Reverse transcription–polymerase chain reaction

SDS:

Sodium dodecyl sulfate

TRPV4:

Transient receptor potential vanilloid 4

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Acknowledgements

The authors acknowledge the assistance of the Research Equipment Sharing Center at Nagoya City University.

Funding

This work was supported in part by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science, KAKEN grant numbers 16K10101, 17K10197, 18K07832, and 21J13766. This work was also supported by a Grant-in-Aid for Research in Nagoya City University, grant numbers 2214008 and 2222003.

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Author notes

  1. Naoya Fukuda and Kohki Toriuchi have contributed equally to this work.

Authors and Affiliations

  1. Department of Pathobiology, Nagoya City University Graduate School of Pharmaceutical Sciences, 3-1 Tanabedori, Mizoho-Ku, Nagoya, Aichi, 467-8603, Japan

    Naoya Fukuda, Kohki Toriuchi, Rina Mimoto, Hiromasa Aoki, Hiroki Kakita, Satoru Takeshita & Mineyoshi Aoyama

  2. Department of Perinatal and Neonatal Medicine, Aichi Medical University, 1-1 Yazakokarimata, Nagakute, Aichi, 480-1195, Japan

    Hiroki Kakita, Satoru Takeshita & Yasumasa Yamada

  3. Department of Molecular and Cellular Pharmacology, Nagoya City University Graduate School of Pharmaceutical Sciences, 3-1 Tanabedori, Mizoho-Ku, Nagoya, Aichi, 467-8603, Japan

    Yoshiaki Suzuki & Hisao Yamamura

  4. Department of Anesthesiology and Intensive Care Medicine, Nagoya City University Graduate School of Medical Sciences, 1 Kawasumi, Mizuho-Ku, Nagoya, Aichi, 467-8601, Japan

    Tetsuya Tamura

  5. Department of Cell Signaling, Nagoya City University Graduate School of Pharmaceutical Sciences, 3-1 Tanabe-Dori, Mizuho-Ku, Nagoya, Aichi, 467-8603, Japan

    Yasumichi Inoue & Hidetoshi Hayashi

  6. Department of Innovative Therapeutic Sciences, Cooperative Major in Nanopharmaceutical Sciences, Nagoya City University Graduate School of Pharmaceutical Sciences, 3-1 Tanabe-Dori, Mizuho-Ku, Nagoya, Aichi, 467-8603, Japan

    Yasumichi Inoue & Hidetoshi Hayashi

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  1. Naoya Fukuda
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Contributions

NF and KT designed this study, preformed all experiments, analyzed and interpreted the data, and wrote the manuscript. RM, HA, HK, and YS collected and assembled the data, performed data analysis and interpretation, and wrote the manuscript. ST, TT, HY, YI, HH, and YY collected and assembled the data and performed data analysis and interpretation. MA designed the study, analyzed and interpreted the data, and wrote the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Mineyoshi Aoyama.

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The authors declare that they have no competing interests.

Data Availability

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Ethical Approval

The present study was approved by the Animal Care and Use Committee of Nagoya City University Graduate School of Pharmaceutical Sciences (protocol number, H27-P-03), and all experiments were performed in accordance with institutional and U.S. National Institutes of Health guidelines for the care and use of laboratory animals.

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Fukuda, N., Toriuchi, K., Mimoto, R. et al. Hypothermia Attenuates Neurotoxic Microglial Activation via TRPV4. Neurochem Res 49, 800–813 (2024). https://doi.org/10.1007/s11064-023-04075-8

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