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METTL3 Mediates Microglial Activation and Blood–Brain Barrier Permeability in Cerebral Ischemic Stroke by Regulating NLRP3 Inflammasomes Through m6A Methylation Modification

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Abstract

Cerebral ischemic stroke (CIS) is the main cause of disability. METTL3 is implicated in CIS, and we explored its specific mechanism. Middle cerebral artery occlusion (MCAO) rat model and oxygen–glucose deprivation/reperfusion (OGD/R) HAPI cell model were established and treated with LV-METTL3 or DAA, oe-METTL3, miR-335-3p mimics, or DAA, to assess their effects on MCAO rat neurological and motor function, cerebral infarction area, brain water content, microglial activation, blood–brain barrier (BBB) permeability, and NLRP3 inflammasome activation. METTL3, pri-miR-335-3p, mature miR-335-3p, and miR-335-3p mRNA levels were assessed by RT-qPCR; M1/M2 microglial phenotype proportion and M1/M2 microglia ratio, inflammatory factor levels, and m6A modification were assessed. MCAO rats manifested cerebral ischemia injury. METTL3 was under-expressed in CIS. METTL3 overexpression inhibited microglial activation and M1 polarization and BBB permeability in MCAO rats and inhibited OGD/R-induced microglial activation and reduced M1 polarization. METTL3 regulated miR-335-3p expression and inhibited NLRP3 inflammasome activation. m6A methylation inhibition averted METTL3’s effects on NLRP3 activation, thus promoting microglial activation in OGD/R-induced cells and METTL3’s effects on BBB permeability in MCAO rats. Briefly, METTL3 regulated miR-335-3p expression through RNA m6A methylation and inhibited NLRP3 inflammasome activation, thus repressing microglial activation, BBB permeability, and protecting against CIS.

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The data that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This work was supported by the Liaoning Provincial Department of Education Research of Science and Technology Youth Talent “Seedling” Project (effect of post-ischemic adaptation of distal limb on the prognosis of acute cerebral infarction and the effect of clinical staging) (NO.JYTQN2020035).

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Authors and Affiliations

  1. Department of Clinical Nutrition, The First Affiliated Hospital of Jinzhou Medical University, Jinzhou, 121000, China

    Xue Cheng

  2. Clinical Medicine, The First Clinical Medical College, Jinzhou Medical University, Jinzhou, 121000, China

    Zhetan Ren

  3. Neurology, Jinzhou Medical University, Jinzhou, 121000, China

    Huiyang Jia

  4. Department of Tumor Intervention, The First Affiliated Hospital of Jinzhou Medical University, No. 2, Section 5, Renmin Street, Guta District, Jinzhou, 121000, China

    Gang Wang

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Contributions

X.C. is the guarantor of integrity of the entire study and contributed to the literature research, manuscript preparation, and editing; X.C. and G.W. contributed to the study concepts, study design, and data analysis; G.W. contributed to the definition of intellectual content and manuscript review; X.C., Z.T.R., and H.Y.J. contributed to the data acquisition; X.C. and Z.T.R. contributed to the experimental studies; X.C. and H.Y.J. contributed to the statistical analysis.

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Correspondence to Gang Wang.

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Cheng, X., Ren, Z., Jia, H. et al. METTL3 Mediates Microglial Activation and Blood–Brain Barrier Permeability in Cerebral Ischemic Stroke by Regulating NLRP3 Inflammasomes Through m6A Methylation Modification. Neurotox Res 42, 15 (2024). https://doi.org/10.1007/s12640-024-00687-2

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