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Pretreatment with Clodronate Improved Neurological Function by Preventing Reduction of Posthemorrhagic Cerebral Blood Flow in Experimental Subarachnoid Hemorrhage

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Abstract

Background

Brain perivascular macrophages (PVMs) are potential treatment targets for subarachnoid hemorrhage (SAH), and previous studies revealed that their depletion by clodronate (CLD) improved outcomes after experimental SAH. However, the underlying mechanisms are not well understood. Therefore, we investigated whether reducing PVMs by CLD pretreatment improves SAH prognosis by inhibiting posthemorrhagic impairment of cerebral blood flow (CBF).

Methods

In total, 80 male Sprague–Dawley rats received an intracerebroventricular injection of the vehicle (liposomes) or CLD. Subsequently, the rats were categorized into the prechiasmatic saline injection (sham) and blood injection (SAH) groups after 72 h. We assessed its effects on weak and severe SAH, which were induced by 200- and 300-µL arterial blood injections, respectively. In addition, neurological function at 72 h and CBF changes from before the intervention to 5 min after were assessed in rats after sham/SAH induction as the primary and secondary end points, respectively.

Results

CLD significantly reduced PVMs before SAH induction. Although pretreatment with CLD in the weak SAH group provided no additive effects on the primary end point, rats in the severe SAH group showed significant improvement in the rotarod test. In the severe SAH group, CLD inhibited acute reduction of CBF and tended to decrease hypoxia-inducible factor 1α expression. Furthermore, CLD reduced the number of PVMs in rats subjected to sham and SAH surgery, although no effects were observed in oxidative stress and inflammation.

Conclusions

Our study proposes that pretreatment with CLD-targeting PVMs can improve the prognosis of severe SAH through a candidate mechanism of inhibition of posthemorrhagic CBF reduction.

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Data Availability

The original contributions presented in the study are included in the article; further inquiries can be directed to the corresponding author.

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Acknowledgements

We would like to thank Editage (www.editage.com) for the English language editing.

Funding

This study was funded by JSPS KAKENHI (grant 19K09459).

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Authors and Affiliations

  1. Department of Pharmacology and Molecular Therapeutics, Kumamoto University Graduate School of Medical Sciences, Kumamoto, Japan

    Hiroki Uchikawa, Koki Kameno & Shokei Kim-Mitsuyama

  2. Department of Neurosurgery, Kumamoto University Graduate School of Medical Sciences, Kumamoto, Japan

    Hiroki Uchikawa, Koki Kameno, Keitaro Kai, Ken Uekawa & Akitake Mukasa

  3. Department of Neurosurgery, Kurume University School of Medicine, Kurume, Japan

    Sosho Kajiwara, Kana Fujimori & Yu Hasegawa

  4. Department of Cell Pathology, Kumamoto University Graduate School of Medical Sciences, Kumamoto, Japan

    Yukio Fujiwara

  5. Department of Pharmaceutical Science, School of Pharmacy at Fukuoka, International University of Health and Welfare, Okawa, Japan

    Yu Hasegawa

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Contributions

HU and YH contributed to the study’s conception and design. HU, K. Kameno, K. Kai, SK, KF, KU, and YF performed the experiments. HU and YH performed the statistical analysis. AM and SK-M helped with the interpretations. HU wrote the first draft of the manuscript. YH revised the manuscript. All authors reviewed and approved the manuscript.

Corresponding author

Correspondence to Yu Hasegawa.

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The authors declare that they have no conflicts of interest.

Ethical Approval

All experiments were approved by the Institutional Animal Care and Use Committee of Kumamoto University, and all applicable institutional guidelines for the care and use of animals were followed.

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Uchikawa, H., Kameno, K., Kai, K. et al. Pretreatment with Clodronate Improved Neurological Function by Preventing Reduction of Posthemorrhagic Cerebral Blood Flow in Experimental Subarachnoid Hemorrhage. Neurocrit Care 39, 207–217 (2023). https://doi.org/10.1007/s12028-023-01754-w

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