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Cited by 34Extracellular vesicles in diseaseBarrier Structure and Function StudiesNeuroinflammation and Neurodegeneration Mechanisms

M2 Microglial Extracellular Vesicles Attenuated Blood-brain Barrier Disruption via MiR-23a-5p in Cerebral Ischemic Mice

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Jiaji Pan, Lin Qi, Liping Wang, Chang Liu, Yaying Song, Muyassar Mamtilahun, Xiaowen Hu, Yong-Fang Li +7 more·2023·aging and disease
DOI 10.14336/ad.2023.0714PMC PMC11081152
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Abstract

Protecting the integrity of the blood-brain barrier (BBB) is crucial for maintaining brain homeostasis after ischemic stroke. Previous studies showed that M2 microglial extracellular vesicles (EVs) played a neuroprotective role in cerebral ischemia. However, the role of M2 microglial EVs in maintaining BBB integrity is unclear. Therefore, we explored the mechanisms of M2 microglial EVs in regulating BBB integrity. To identify microglial EVs, we used nanoparticle tracking analysis, transmission electron microscopy, and western blot analysis. Adult male ICR mice were subjected to 90-min middle cerebral artery occlusion (MCAO), followed by the injection of PKH26-labeled M2 microglial EVs via the tail vein. After MCAO, we assessed brain infarct and edema volume, as well as modified neurological severity score. BBB integrity was measured by assessing IgG leakage. The effects of M2 microglial EVs on astrocytes and endothelial cells were also examined. To investigate the molecular mechanisms, we performed RNA sequencing, miR-23a-5p knockdown, and luciferase reporter assays. Our results showed that PKH26-labeled microglial EVs were mainly taken up by neurons and glial cells. M2 microglial EVs treatment decreased brain infarct and edema volume, modified neurological severity score, and IgG leakage, while increasing the ZO-1, occludin, and claudin-5 expression after MCAO. Knockdown of miR-23a-5p reversed these effects. RNA sequencing revealed that the TNF, MMP3 and NFκB signaling pathway involved in regulating BBB integrity. Luciferase reporter assay showed that miR-23a-5p could bind to the 3' UTR of TNF. M2 microglial EVs-derived miR-23a-5p decreased TNF, MMP3 and NFκB p65 expression in astrocytes after oxygen-glucose deprivation, thereby increasing ZO-1 and Claudin-5 expression in bEnd.3 cells. In conclusion, our findings demonstrated that M2 microglial EVs attenuated BBB disruption after cerebral ischemia by delivering miR-23a-5p, which targeted TNF and regulated MMP3 and NFκB p65 expression.

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  • Vesicle-derived miR-23a-5p can directly target TNF, by down-regulating MMP3 and NF-κB p65 to regulate the integrity of hemoglobin and attenuate destruction of the blood-brain barrier after cerebral ischemia.

    “The vesicle-derived miR-23a-5p could directly target TNF, by down-regulating the expression of MMP3 and NF-κB p65 to regulate the integrity of hemoglobin and attenuates the destruction of the blood-brain barrier after cerebral ischemia [5].”

    From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [5]

    The Results section reports that vesicle-derived miR-23a-5p binds the TNF 3′-UTR, reduces TNF, MMP3 and NFκB p65 expression, and helps maintain blood–brain barrier integrity in ischemic mice. It assesses IgG leakage and tight-junction proteins, not the integrity of hemoglobin; that part of the citing sentence is unsupported. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • Treatment with M2 microglia-derived extracellular vesicles could reduce the area of cerebral infarction and edema in mice.

    “Moreover, the experiment in mice confirmed that the treatment with M2 microglia-derived extracellular vesicles could reduce the area of cerebral infarction and edema.”

    From Recent advances of miR-23 in human diseases and growth development (2025) · cites this paper as [5]

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Authors
Pan, J., Qi, L., Wang, L., Liu, C., Song, Y., Mamtilahun, M., Hu, X., Li, Y.-F., Chen, X., Khan, H. L., Xu, Q., Wang, Y., Tang, Y., Yang, G., & Zhang, Z.
Affiliations
Shanghai Jiao Tong University (CN)
Corresponding author
Zhijun Zhang
ORCID
Venue
aging and disease
Publisher
Aging and Disease
Published
2023
Volume
15
Issue
3
Pages
1344-1356
ISSN
2152-5250 (ISSN-L)
DOI
10.14336/ad.2023.0714
PMC
PMC11081152
OpenAlex
W4386117009
Cited by
34
References
33
Citations by year
2026: 4 · 2025: 17 · 2024: 11 · 2023: 2
FWCI
3.03
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Aging and Disease (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Natural Science Foundation of China (82071284, 82271320, 81974179, 2019-01)
  • Science and Technology Commission of Shanghai Municipality
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Life Sciences › Neuroscience › Neurology › Barrier Structure and Function Studies
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
M2 microglia, extracellular vesicles, blood-brain barrier integrity, miR-23a-5p, cerebral ischemia, ischemic stroke, middle cerebral artery occlusion, TNF-α, MMP-3, NF-κB p65, tight junction proteins, ZO-1, claudin-5, astrocytes, neuroprotection, microRNA delivery
MeSH terms
Extracellular Vesicles, Animals, Astrocytes, Blood-Brain Barrier, Brain Ischemia, Disease Models, Animal, Male, Mice, Inbred ICR, Microglia, Matrix Metalloproteinase 3, Infarction, Middle Cerebral Artery, MicroRNAs, Mice, Transcription Factor RelA, Claudin-5, Zonula Occludens-1 Protein

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