Sevoflurane Post-Conditioning Ameliorates Neuronal Deficits and Axon Demyelination After Neonatal Hypoxic Ischemic Brain Injury: Role of Microglia/Macrophage

Abstract

Microglia/macrophages have been identified to be highly polarized after ischemia. Interestingly, the polarization of these microglia/macrophages varies immensely under differing disease conditions. Post-conditioning using sevoflurane, a volatile anesthetic, could provide long-term neuroprotection to neonatal rats after hypoxic-ischemic brain injury (HIBI). Thus, the current study aimed at investigating the effects of sevoflurane post-conditioning (SPC) on microglia/macrophage polarization after HIBI induction in neonatal rats. Additionally, we aimed at identifying the underpinning mechanisms specifically related to autophagy and lysosomal protease enzyme, cathepsin B. To develop a HIBI model, 7-day-old Sprague–Dawley rats underwent left common carotid artery ligation followed by 2 h of hypoxia. The role of microglia/macrophages in the neuroprotection conferred by SPC was examined by left-side intra-cerebroventricular injection with adenovirus vector carrying catB-GFP or rapamycin. The number of interleukin (IL)-1β+ cells, cathepsin B+ cells, light chain 3B positive (LC3B+) cells among ionized calcium binding adaptor molecule 1(Iba1+)cells to investigate microglia polarization, neuronal apoptosis to assess neuronal death in the acute phase were tested at 24 h after HIBI. Behavioral tests including suspension test, Morris water maze tests were performed to investigate the long-term effects of SPC, at 21 to 34 days post HIBI. Nissl staining and myelin basic protein (MBP) immunostaining to assess the long-term neuronal and myelin damage were performed at 34 days after HIBI. Based on the obtained results post HIBI, we observed the cells that were positive for IL-1β, cathepsin B, and LC3B among Iba1 positive cell population in the hippocampus were significantly decreased after SPC treatment. SPC significantly attenuated the HIBI-induced increase in neuronal apoptosis, improved long-term cognitive function, and attenuated HI-induced decrease of Nissl-positive cells and MBP expression. However, these trends were reversed by injection of adenovirus vector carrying catB-GFP and rapamycin. SPC attenuated microglia polarization towards neurotoxic phenotypes, alleviates neuronal death and axon demyelination after HIBI in neonatal rats by regulating microglia autophagy and cathepsin B expression, and therefore provided long-term cognitive, learning and memory protection.

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Abbreviations

HIBI:

Hypoxic-ischemic brain injury

HI:

Hypoxia ischemia

SPC:

Sevoflurane post-conditioning

CA:

Cornu ammonis

MWM:

Morris water maze

DG:

Dentate gyrus

CNS:

Central nervous system

iNOS:

Induced nitrogen monoxide synthase

DAB:

Diaminobenzidine

1Iba1:

Ionized calcium binding adaptor molecule

MBP:

Myelin basic protein

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Acknowledgements

Thanks to Prof. Lili Wang’s suggestion and help in completing the experiments.

Funding

This research is supported by grant from the National Natural Science Foundation of China (Nos. 81671311, 81870838), Liaoning Province Distinguished Professor Support Program (No. XLYC1802096) and Outstanding Scientific Fund of Shengjing Hospital (No. 201708), and Colleges and Universities Basic Research Project Fund of Liaoning Province (No. LQNK201709).

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Contributions

H.X. did the experiment and wrote the paper; Q.G. and J.N. contributed in statistical analysis; Y.Z., Z.W. and C.L. did the experiment; Ping Zhao designed the experiment and offered the fund.

Corresponding author

Correspondence to Ping Zhao.

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The authors declare that they have no conflict of interest.

Ethical Approval

Animal experiments were all performed according to the recommendations of the National Institutes of Health Guidelines for the Care and Use of Laboratory animals. Protocols described below were all approved by the Animal Review Board of Shengjing Hospital, China Medical University. This article does not contain any studies with human participants performed by any of the authors.

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Xue, H., Zhang, YH., Gao, QS. et al. Sevoflurane Post-Conditioning Ameliorates Neuronal Deficits and Axon Demyelination After Neonatal Hypoxic Ischemic Brain Injury: Role of Microglia/Macrophage. Cell Mol Neurobiol (2020). https://doi.org/10.1007/s10571-020-00949-5

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Keywords

  • Sevoflurane post-conditioning
  • Hypoxic-ischemic brain injury
  • Neonatal rat
  • Microglia/macrophage
  • Cathepsin B