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Gas6 Promotes Microglia Efferocytosis and Suppresses Inflammation Through Activating Axl/Rac1 Signaling in Subarachnoid Hemorrhage Mice

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An Author Correction to this article was published on 08 April 2024

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Abstract

Early brain injury (EBI) following  subarachnoid hemorrhage (SAH) is characterized by rapid development of neuron apoptosis and dysregulated inflammatory response. Microglia efferocytosis plays a critical role in the clearance of apoptotic cells, attenuation of inflammation, and minimizing brain injury in various pathological conditions. Here, using a mouse SAH model, we aim to investigate whether microglia efferocytosis is involved in post-SAH inflammation and to determine the underlying signaling pathway. We hypothesized that TAM receptors and their ligands regulate this process. To prove our hypothesis, the expression and cellular location of TAM (Tyro3, Axl, and Mertk) receptors and their ligands growth arrest-specific 6 (Gas6) and Protein S (ProS1) were examined by PCR, western blots, and fluorescence immunostaining. Thirty minutes after SAH, mice received an intraventricular injection of recombinant Gas6 (rGas6) or recombinant ProS1 (rPros1) and underwent evaluations of inflammatory mediator expression, neurological deficits, and blood–brain barrier integrity at 24 h. Microglia efferocytosis of apoptotic neurons was analyzed in vivo and in vitro. The potential mechanism was determined by inhibiting or knocking down TAM receptors and Rac1 by specific inhibitors or siRNA. SAH induced upregulation of Axl and its ligand Gas6. The administration of rGas6 but not rPros1 promoted microglia efferocytosis, alleviated inflammation, and ameliorated SAH-induced BBB breakdown and neurological deficits. The beneficial effects of rGas6 were arrogated by inhibiting or knocking down Axl and Rac1. We concluded that rGas6 attenuated the development of early brain injury in mice after SAH by facilitating microglia efferocytosis and preventing inflammatory response, which is partly dependent on activation of Axl and Rac1.

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The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.

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Funding

This research was supported by grants from the National Natural Science Foundation of China No. 82071283 to Qin Hu, No. 81601013 to Junjia Tang, No. 31900686 to Jianfei Lu, and No. 82271310 to Feng Jia, the Natural Science Foundation of Shanghai 22ZR1437700 to Qin Hu, and Science and Technology Commission of Shanghai Municipal 21ZR1439000 to Feng Jia.

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Author notes

  1. Junjia Tang, Yichao Jin and Feng Jia contributed equally to this work and shared the first authorship.

Authors and Affiliations

  1. Department of Neurosurgery, Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200127, China

    Junjia Tang, Yichao Jin, Feng Jia, Tao Lv, Lin-Feng Zhang, Zeyu Zhang, Bin Zhao, Xiaohua Zhang & Qin Hu

  2. Department of Neurosurgery, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 201620, China

    Junjia Tang & Yajun Xue

  3. Department of Neurology, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Anatol Manaenko

  4. Discipline of Neuroscience, Department of Physiology and Anatomy, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China

    Xin Qi & Jianfei Lu

  5. Department of Physiology and Pharmacology, Loma Linda University, Loma Linda, CA, USA

    John H. Zhang

  6. Songjiang Institute and Songjiang Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 201600, China

    Jianfei Lu

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  1. Junjia Tang
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Contributions

JT, JL, YJ, and QH conceived and designed the study. JT and TL performed animal models and behavioral tests, and L-FZ and XQ did the in vitro microglia efferocytosis experiment. YJ and ZZ collected the samples and run PCR and western blots. JL, BZ, and JT performed the histology and immunostaining. JL, XZ, and QH analyzed the data and finalized the figures. FJ, AM, JHZ, and QH wrote the manuscript. All the authors revised and approved the final version of the manuscript.

Corresponding authors

Correspondence to Jianfei Lu or Qin Hu.

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Ethics Approval and Consent to Participate

All experimental procedures involving animals were approved by the Ren Ji Hospital Animal Care and Use Committee and were performed in accordance with the guidelines in the National Institutes of Health (NIH) Guide for the Care and Use of Laboratory Animals and followed the ARRIVE guidelines. This article does not contain any studies with human participants performed by any of the authors.

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Tang, J., Jin, Y., Jia, F. et al. Gas6 Promotes Microglia Efferocytosis and Suppresses Inflammation Through Activating Axl/Rac1 Signaling in Subarachnoid Hemorrhage Mice. Transl. Stroke Res. 14, 955–969 (2023). https://doi.org/10.1007/s12975-022-01099-0

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