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Effects of Toll-Like Receptor 4 Antagonists Against Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage in Mice

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Abstract

Toll-like receptor 4 (TLR4) signaling may play a crucial role in the occurrence of cerebral vasospasm after subarachnoid hemorrhage (SAH). The main purpose of this study was to assess if selective blockage of TLR4 on cerebral arteries prevents cerebral vasospasm development and neurological impairments after SAH in mice. One hundred fourteen mice underwent endovascular perforation SAH or sham operation and were randomly divided into the following 6 groups: sham+vehicle, sham+LPS-RS ultrapure 8 μg, sham+LPS-RS ultrapure 40 μg, SAH+vehicle, SAH+LPS-RS ultrapure 8 μg, and SAH+LPS-RS ultrapure 40 μg. A selective TLR4 antagonist, LPS-RS ultrapure (8 or 40 μg), was administered intracerebroventricularly to mice at 30 min, and the effects were evaluated by neurobehavioral tests and India-ink angiography at 24–48 h, and Western blotting and immunohistochemistry on cerebral arteries at 24 h post-SAH. Higher but not lower dosages of LPS-RS ultrapure significantly prevented post-SAH neurological impairments and ameliorated cerebral vasospasm. SAH caused TLR4 activation and cyclooxygenase-1 (COX1) upregulation in the endothelial cells and smooth muscle cells of spastic cerebral arteries, both of which were significantly suppressed by LPS-RS ultrapure. Another selective TLR4 antagonist, IAXO-102, which has a different binding site from LPS-RS ultrapure, also showed similar protective effects to LPS-RS ultrapure. These findings suggest that TLR4 signaling is implicated in cerebral vasospasm development at least partly via COX1 upregulation, and that TLR4 antagonists have therapeutic potential as a new therapy against cerebral vasospasm.

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Abbreviations

ACA:

Anterior cerebral artery

ANOVA:

Analysis of variance

BA:

Basilar artery

COX:

Cyclooxygenase

COX1:

Cyclooxygenase-1

COX2:

Cyclooxygenase-2

CSF:

Cerebrospinal fluid

DCI:

Delayed cerebral ischemia

ET:

Endothelin

HE:

Hematoxylin and eosin

ICA:

Internal carotid artery

ICAM:

Intercellular adhesion molecule

IL:

Interleukin

MCA:

Middle cerebral artery

MCP:

Monocyte chemoattractant protein

NO:

Nitric oxide

PBS:

Phosphate-buffered saline

NF:

Nuclear factor

SAH:

Subarachnoid hemorrhage

TLR4:

Toll-like receptor 4

TNF:

Tumor necrosis factor

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Acknowledgments

We thank Ms. Chiduru Yamamoto (Department of Neurosurgery, Mie University Graduate School of Medicine) for her technical assistance. This work was supported by a Grant-in-Aid for Scientific Research from SENSHIN Medical Research Foundation to Dr. Suzuki, and Japan Society for the Promotion of Science to Dr. Fujimoto.

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Authors and Affiliations

  1. Department of Neurosurgery, Mie University Graduate School of Medicine, 2-174 Edobashi, Tsu, Mie, 514-8507, Japan

    Fumihiro Kawakita, Masashi Fujimoto, Lei Liu, Fumi Nakano, Yoshinari Nakatsuka & Hidenori Suzuki

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  1. Fumihiro Kawakita
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  2. Masashi Fujimoto
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  3. Lei Liu
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  4. Fumi Nakano
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  5. Yoshinari Nakatsuka
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  6. Hidenori Suzuki
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Corresponding author

Correspondence to Hidenori Suzuki.

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Funding

This work was funded by a Grant-in-Aid for Scientific Research from SENSHIN Medical Research Foundation to Dr. Suzuki, and Japan Society for the Promotion of Science to Dr. Fujimoto.

Conflict of Interest

The authors declare that they have no conflict of interest.

Research Involving Animals

All procedures were approved by the Animal Ethics Review Committee of Mie University and were carried out according to the institution’s Guidelines for Animal Experiments.

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Kawakita, F., Fujimoto, M., Liu, L. et al. Effects of Toll-Like Receptor 4 Antagonists Against Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage in Mice. Mol Neurobiol 54, 6624–6633 (2017). https://doi.org/10.1007/s12035-016-0178-7

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