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Updated: Jan 9, 2026
Isolation of Cortical Microglia with Preserved Immunophenotype and Functionality From Murine Neonates
Published on: January 30, 2014
Methylnissolin mitigates microglia-mediated neuroinflammation and ischemic brain injury through PI3K/AKT and MAPK
Jie Wu1, Maoyuan Jiang1, Yanping Yin1
1Department of Neurology, Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine, Nanjing, Jiangsu, 210008, China; Department of Neurology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Background:
Neuroinflammation driven by activated microglia is a major contributor to secondary brain damage following ischemic stroke. Targeting microglial activation has thus emerged as an important therapeutic strategy. Methylnissolin (ML), a pterocarpan-type isoflavonoid with documented anti-inflammatory activity and predicted ability to cross the blood-brain barrier, represents a potential modulator of microglial responses.
Methods:
Primary microglia subjected to oxygen-glucose deprivation/reoxygenation (OGD/R) and mice subjected to transient middle cerebral artery occlusion (tMCAO) were treated with ML. Microglial inflammatory responses were measured by RT-qPCR, Western blotting, Enzyme-linked immunosorbent assay, and immunofluorescence. Brain pathological alterations were assessed by 2,3,5-triphenyltetrazolium chloride (TTC) staining, Evans blue extravasation, and brain water content measurement. Behavioral changes was evaluated using grip strength, rotarod, modified neurological severity score (mNSS), and foot-fault tests. RNA sequencing (RNA-seq) and targeted protein-level analyses were performed to investigate signaling pathways.
Results:
ML markedly reduced the OGD/R-induced expression of pro-inflammatory mediators (IL-1β, IL-6, TNF-α, iNOS, COX-2) and suppressed activation of the PI3K/AKT and MAPK pathways in microglia. In tMCAO mice, ML treatment lessened infarct volume, brain edema, and blood-brain barrier leakage, preserved neuronal morphology, and improved neurological performance. RNA sequencing and biochemical analyses in primary microglia showed modulation of the PI3K/AKT and MAPK signaling cascades by ML.
Conclusion:
ML mitigates microglia-mediated neuroinflammation and protects against ischemic brain injury by downregulating the PI3K/AKT and MAPK pathways, suggesting that ML may be a promising therapeutic candidate for ischemic stroke.
Insights
Methylnissolin (ML) reduces neuroinflammation and brain damage after ischemic stroke by suppressing microglial activation. This compound shows promise as a novel therapeutic for stroke by modulating key inflammatory pathways.
Area of Science:
- Neuroscience
- Pharmacology
- Molecular Biology
Background:
- Microglial activation drives secondary brain damage in ischemic stroke.
- Targeting microglial pathways is a key therapeutic strategy.
- Methylnissolin (ML), a blood-brain barrier-permeable isoflavonoid, may modulate microglial responses.
Purpose of the Study:
- To investigate the therapeutic potential of Methylnissolin (ML) in mitigating ischemic brain injury.
- To elucidate the effects of ML on microglial activation and associated inflammatory pathways.
Main Methods:
- Primary microglia and mice underwent oxygen-glucose deprivation/reoxygenation (OGD/R) or transient middle cerebral artery occlusion (tMCAO) and were treated with ML.
- Microglial inflammatory markers, brain pathology, and neurological function were assessed.
- RNA sequencing and protein analyses identified affected signaling pathways.
Main Results:
- ML suppressed pro-inflammatory mediators (IL-1β, IL-6, TNF-α) and PI3K/AKT/MAPK pathway activation in microglia.
- ML reduced infarct volume, brain edema, and blood-brain barrier leakage in stroke models.
- ML improved neurological performance and preserved neuronal morphology.
Conclusions:
- ML effectively mitigates microglia-driven neuroinflammation and ischemic brain injury.
- The protective effects of ML are mediated by the downregulation of PI3K/AKT and MAPK signaling.
- ML represents a potential therapeutic candidate for treating ischemic stroke.
