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Updated: May 28, 2025
Characterization and Isolation of Mouse Primary Microglia by Density Gradient Centrifugation
Published on: February 16, 2018
Catalpalactone protects rats nerve function from hypoxic lesion by polarizing microglial cells toward M2 phenotype
1Department of Neurology Ward 2, The Third Affiliated Hospital, Qiqihar Medical University, Qiqihar, 161000, China. aminda001@qmu.edu.cn.
Background:
Ischemic brain injury results in high disability due to neuroinflammation and oxidative stress, and M1/M2 polarization of glial cells plays a key role in neuroinflammation. This research explored the protective effect of Catalpalactone on middle cerebral artery occlusion (MCAO)-induced brain injury and its underlying regulation mechanism in rats.
Methods:
The ischemic lesions were induced by the MCAO, and the oxygen and glucose deprivation/reoxygenation (OGD/R) was used for BV2 microglial cell induction. The polarization of glial cells was determined via immunohistochemistry staining assessment. Oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) assays were used for the glycolysis and oxidative phosphorylation test. After that, the cell counting kit-8 (CCK-8) for cell viability test and flow cytometry for apoptosis and phosphorylation analysis were performed. Furthermore, a co-culture model of BV2 and PC12 cells was used for the purpose of exploring the effects of Catalpalactone on the interaction and of microglia and neurons in ischemic brain injury. Finally, the Modified Neurological Severity Score (mNSS) analysis was used for the analysis on the neurological function.
Results:
After MCAO induction, the infiltration of microglial cells were significantly increased in the injury area, and its M1 phenotype was enhanced (up-regulated Cd86). In vitro, the OGD/R-induced BV2 microglial cell also exhibited the increasing M1 phenotype with higher glycolysis activity, but lower oxidative phosphorylation through the activating JAK-SATA signaling pathway. Finally, we determined that 15 μM Catalpalactone optimally induces M2 microglial polarization with increased cell viability and decreased apoptosis in the OGD/R-induced BV2 cell model, while also reducing mNSS scores and improving neurological function in the MCAO rat model.
Conclusion:
We clarified the underlying mechanism of Catalpalactone treatment for ischemic lesions through promoting M2 microglial cells phenotype.
Insights
Catalpalactone protects against ischemic brain injury by promoting M2 glial cell polarization. This neuroprotective effect reduces inflammation and improves neurological function following middle cerebral artery occlusion in rats.
Area of Science:
- Neuroscience
- Biochemistry
- Pharmacology
Background:
- Ischemic brain injury causes significant disability via neuroinflammation and oxidative stress.
- Glial cell polarization, specifically M1/M2 phenotypes, is a critical factor in neuroinflammation.
- Understanding these mechanisms is key to developing effective treatments for stroke.
Purpose of the Study:
- To investigate the protective effects of Catalpalactone on middle cerebral artery occlusion (MCAO)-induced brain injury in rats.
- To elucidate the underlying regulatory mechanisms of Catalpalactone, focusing on glial cell polarization.
- To assess Catalpalactone's impact on neuroinflammation and neuronal survival.
Main Methods:
- Middle cerebral artery occlusion (MCAO) model in rats and oxygen-glucose deprivation/reoxygenation (OGD/R) in BV2 microglial cells.
- Immunohistochemistry for glial cell polarization, OCR/ECAR assays for metabolic activity, CCK-8 for cell viability, and flow cytometry for apoptosis.
- Co-culture models and Modified Neurological Severity Score (mNSS) for functional assessment.
Main Results:
- MCAO induced significant microglial infiltration and M1 phenotype (Cd86 upregulation) in the injured brain area.
- OGD/R-treated BV2 cells showed M1 polarization, increased glycolysis, and reduced oxidative phosphorylation via JAK-STAT signaling.
- 15 μM Catalpalactone promoted M2 polarization, enhanced cell viability, reduced apoptosis in vitro, and improved neurological function in vivo.
Conclusions:
- Catalpalactone treatment effectively promotes M2 microglial cell polarization.
- This shift in glial cell phenotype underlies Catalpalactone's therapeutic mechanism for ischemic brain lesions.
- Catalpalactone demonstrates significant neuroprotective potential in ischemic stroke models.
