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IRAK-M Ablation Promotes Status Epilepticus-Induced Neuroinflammation via Activating M1 Microglia and Impairing Excitatory Synaptic Function

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Abstract

Epilepsy is one of the most common neurological disorders. The pro-epileptic and antiepileptic roles of microglia have recently garnered significant attention. Interleukin-1 receptor-associated kinase (IRAK)-M, an important kinase in the innate immune response, is mainly expressed in microglia and acts as a negative regulator of the TLR4 signaling pathway that mediates the anti-inflammatory effect. However, whether IRAK-M exerts a protective role in epileptogenesis as well as the molecular and cellular mechanisms underlying these processes are yet to be elucidated. An epilepsy mouse model induced by pilocarpine was used in this study. Real-time quantitative polymerase chain reaction and western blot analysis were used to analyze mRNA and protein expression levels, respectively. Whole-cell voltage-clamp recordings were employed to evaluate the glutamatergic synaptic transmission in hippocampal neurons. Immunofluorescence was utilized to show the glial cell activation and neuronal loss. Furthermore, the proportion of microglia was analyzed using flow cytometry. Seizure dynamics influenced the expression of IRAK-M. Its knockout dramatically exacerbated the seizures and the pathology in epilepsy and increased the N-methyl-d-aspartate receptor (NMDAR) expression, thereby enhancing glutamatergic synaptic transmission in hippocampal CA1 pyramidal neurons in mice. Furthermore, IRAK-M deficiency augmented hippocampal neuronal loss via a possible mechanism of NMDAR-mediated excitotoxicity. IRAK-M deletion promotes microglia toward the M1 phenotype, which resulted in high levels of proinflammatory cytokines and was accompanied by a visible increase in the expressions of key microglial polarization-related proteins, including p-STAT1, TRAF6, and SOCS1. The findings demonstrate that IRAK-M dysfunction contributes to the progression of epilepsy by increasing M1 microglial polarization and glutamatergic synaptic transmission. This is possibly related to NMDARs, particularly Grin2A and Grin2B, which suggests that IRAK-M could serve as a novel therapeutic target for the direct alleviation of epilepsy.

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Data Availability

Data is contained within the article. The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgments

The authors would like to acknowledge Prof. Hong-Hao Wang from the Department of Neurology, Nanfang Hospital, Southern Medical University for presenting IRAK-M−/− mice, and to express sincere gratitude to the editor and anonymous reviewers for their valuable comments.

Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 81873158, No. 82074265), the Natural Science Foundation of Guangdong Province, China (No.2021A1515011505, No.2020A1515010324), and Construction Fund of Key Disciplines of Traditional Chinese Medicine in Guangdong, China (No. G622299957).

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Authors and Affiliations

  1. School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China

    Xiao-Shan Liang, Ting-Lin Qian, Yi-Fan Xiong, Xiao-Tao Liang, Yue-Wen Ding, Xiao-Yu Zhu, Yun-Lv Li, Jie-Li Zhou, Wei-Peng Li & Wei Xie

  2. Department of Traditional Chinese Medicine, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China

    Yue-Wen Ding & Wei Xie

  3. The First School of Clinical Medicine, Southern Medical University, Guangzhou, 510515, China

    Le-Yi Tan

  4. Department of Neurology, Integrated Hospital of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China

    Wei-Peng Li

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Contributions

Wei Xie, Wei-Peng Li, Yue-Wen Ding, and Xiao-Shan Liang conceived and designed the experiments and revised the manuscript; Ting-Lin Qian, Yi-Fan Xiong, Xiao-Tao Liang, and Xiao-Yu Zhu performed the experiments; Yun-Lv Li, Jie-Li Zhou, and Le-Yi Tan assisted in some of the experimental work; Ting-Lin Qian and Wei-Peng Li analyzed data; The first draft of the manuscript was written by Xiao-Shan Liang, and all authors commented on previous versions of the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Wei-Peng Li or Wei Xie.

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Ethics Approval

All animal experimental protocols in this study were approved by the Southern Medical University Institutional Animal Care and Use Committee (permit number: L2020038) and conducted in accordance with the National Health and Medical Research Council animal ethics and ARRIVE guidelines. The experimental mice were housed in a controlled temperature (23–25°C) and humidity (45–55%) with a modified 12-h dark-light photophase (lights on from 07:00 AM to 07:00 PM) and free access to standard food and water.

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Liang, XS., Qian, TL., Xiong, YF. et al. IRAK-M Ablation Promotes Status Epilepticus-Induced Neuroinflammation via Activating M1 Microglia and Impairing Excitatory Synaptic Function. Mol Neurobiol 60, 5199–5213 (2023). https://doi.org/10.1007/s12035-023-03407-7

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