2. Academic Validation
  3. ANXA1 regulates microglia polarization and autophagy via PI3K/Akt/mTOR pathway to reduce inflammatory injury after cerebral ischemia-reperfusion

ANXA1 regulates microglia polarization and autophagy via PI3K/Akt/mTOR pathway to reduce inflammatory injury after cerebral ischemia-reperfusion

  • Cell Signal. 2026 Mar:139:112336. doi: 10.1016/j.cellsig.2025.112336.
Hua Wang 1,  Xiaohui Li 1,  Ziyu Liu 2,  Ruirui Lu 1,  Xiaopeng Li 1,  Xiaoguang Zhang 3
Affiliations

Affiliations

  • 1 Department of Neurology, the First Affiliated Hospital of Henan University, Kaifeng 475000, Henan Province, China; Institute of Neurological Diseases, Henan University, Kaifeng 475000, Henan Province, China.
  • 2 Department of Neurosurgery, the First Affiliated Hospital of Henan University, Kaifeng 475000, Henan Province, China.
  • 3 Department of Neurosurgery, the First Affiliated Hospital of Henan University, Kaifeng 475000, Henan Province, China. Electronic address: [email protected].
PMID: 41421718 DOI: 10.1016/j.cellsig.2025.112336
Abstract

Background: Annexin A1 (ANXA1) can be activated by ischemia/reperfusion (I/R) events or inflammatory processes, but its specific regulatory mechanisms need further investigation.

Methods: ANXA1 in BV-2 cells was knocked down or overexpressed, and OGD/R induced injury, while 740YP and LY294002 were used for intervention. Microglia polarization phenotype and marker levels were assessed through flow cytometry, RT-qPCR and western blot. Autophagic flux and lysosomal function were evaluated by mCherry-GFP-LC3B, acridine orange (AO) staining and LysoTracker staining. ANXA1 and PI3K/Akt/mTOR proteins were detected by western blot. The tMCAO/R mouse model was established. Longa score, behavioral test and pathological staining to assess the extent of nerve injury, and microglia polarization and Autophagy indicators were detected.

Results: ANXA1 expression was decreased in OGD/R-treated microglia. ANXA1 overexpression facilitated the transformation of microglia phenotype from M1-like phenotype to M2-like phenotype, increased CD206 and IL-10 expression, reduced the GFP/mCherry ratio and LysoTracker positive cells, and increased the red fluorescence of AO staining. ANXA1 overexpression also significantly reduced PI3K, Akt and mTOR phosphorylation. Moreover, ANXA1 overexpression markedly decreased Longa score, brain water content and infarct size, improved motor and neurological impairment in tMCAO/R mice, elevated Nissl bodies and Iba-1+CD206+ positive area, and reduced necrotic neuron numbers, inflammatory factor content and Autophagy protein levels. In addition, 740YP significantly inhibited the improvement of ANXA1 overexpression, and LY294002 significantly enhanced the improvement of ANXA1 overexpression.

Conclusion: Overexpression of ANXA1 regulated microglia polarization and autophagic flux via regulating PI3K/Akt/mTOR pathway, and improved cerebral I/R inflammatory injury.

Keywords

ANXA1; Autophagy; Microglia; PI3K/Akt/mTOR signaling; Polarization.

Figures
Products