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Microglia Activation and Polarization After Intracerebral Hemorrhage in Mice: the Role of Protease-Activated Receptor-1

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Abstract

Polarized microglia play a dual (beneficial/detrimental) role in neurological diseases. However, the status and the factors that modulate microglia polarization in intracerebral hemorrhage (ICH) remain unclear. In the present study, we investigated the role of protease-activated receptor-1 (PAR-1, a thrombin receptor) in ICH-induced microglia polarization in mice. Male wild-type (WT) and PAR-1 knockout (PAR-1 KO) mice received an infusion of 30-μL autologous blood or saline into the right basal ganglia. Mice were euthanized at different time points and the brains were used for Western blotting and immunohistochemistry. Some mice had magnetic resonance imaging. We found that ICH induced microglia activation and polarization. M1 phenotypic markers were markedly increased and reached a peak as early as 4 h, remained high at 3 days and decreased 7 days after ICH. M2 phenotypic markers were upregulated later than M1 markers reaching a peak at day 1 and declining by day 7 after ICH. PAR-1 was upregulated after ICH and expressed in the neurons and microglia. ICH induced less brain swelling and neuronal death in PAR-1 KO mice, and this was associated with less M1 polarization and reduced proinflammatory cytokine levels in the brain. In conclusion, these results suggest that polarized microglia occur dynamically after ICH and that PAR-1 plays a role in the microglia activation and polarization.

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Acknowledgments

This study was supported by grants NS-073595, NS-079157, NS-084049, NS-090925, NS-091545 and NS-096917 from the National Institutes of Health (NIH), 973 Program-2014CB541600, and by grants 81371336, 81400942, 81400945 from National Natural Science Foundation of China (NSFC) and grants LY13H090005 from Natural Science Foundation of Zhejiang Province (ZJNSF). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH, NSFC, and ZJNSF.

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Authors and Affiliations

  1. Department of Neurosurgery, University of Michigan, Ann Arbor, Michigan, USA

    Shu Wan, Yingying Cheng, Hang Jin, Dewei Guo, Ya Hua, Richard F. Keep & Guohua Xi

  2. Department of Neurosurgery, The 1st Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, China

    Shu Wan

  3. Department of Neurology, The 1st Affiliated Hospital, School of Medicine, Jilin University, Changchun, China

    Yingying Cheng & Hang Jin

  4. University of Michigan, Room5018 BSRB, 109 Zina Pitcher Place, Ann Arbor, MI, 48109-2200, USA

    Guohua Xi

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  1. Shu Wan
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Correspondence to Guohua Xi.

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Shu Wan, Yingying Cheng, Hang Jin, Dewei Guo, Ya Hua, Richard F. Keep, and Guohua Xi declare that they have no conflict of interest.

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All institutional and national guidelines for the care and use of laboratory animals were followed.

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ESM 1

Supplemental Figure I: Wild-type (WT) mice received an intracerebral injection of autologous whole blood or saline. At 24 hours, the brains were used to assess PAR-1 immunoreactivity (A) and protein levels (B) in the ipsilateral basal ganglia. Scale bar = 50 μm; values are means ± S.D.; n = 3 per group, *p < 0.01 vs. other groups. Supplemental Figure II: Immunoreactivity of cluster of differentiation 16 (CD16, A) and inducible nitric oxide synthase (iNOS, B) in the ipsilateral basal ganglia 1 day after injection of 30 μL autologous blood or saline into the right caudate. Scale bar=20μm. Supplemental Figure III: Immunoreactivity and protein levels of cluster of differentiation (CD206, A) and chitinase 3 like protein 3 (YM-1, B) in the ipsilateral basal ganglia 1 day after injection of 30 μL autologous blood or saline into the right caudate. Scale bar=20μm. (PDF 28266 kb)

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Wan, S., Cheng, Y., Jin, H. et al. Microglia Activation and Polarization After Intracerebral Hemorrhage in Mice: the Role of Protease-Activated Receptor-1. Transl. Stroke Res. 7, 478–487 (2016). https://doi.org/10.1007/s12975-016-0472-8

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  • DOI: https://doi.org/10.1007/s12975-016-0472-8

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