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Updated: May 11, 2026
Primary Microglia Isolation from Mixed Glial Cell Cultures of Neonatal Rat Brain Tissue
Published on: August 15, 2012
[Gastrodin alleviates microglia-mediated inflammatory responses in neonatal mice with hypoxic-ischemic brain damage
1Department of Human Anatomy & Histology and Embryology, School of Basic Medical Sciences, Kunming Medical University, Kunming 650500, China.
Objective:
To investigate the mechanism behind the protective effects of gastrodin against microglia-mediated inflammatory responses following hypoxic-ischemic brain damage (HIBD) in neonatal mice.
Methods:
Thirty-six 10-day-old C57BL/6J mice were randomized into sham-operated group, HIBD (induced by ligation of the left common carotid artery followed by hypoxia for 40 min) group, and HIBD with gastrodin treatment groups (n=12). In gastrodin treatment group, 100 mg/kg gastrodin was injected intraperitoneally 1 h before and at 2 and 12 h after hypoxia. After the treatments, the expressions of CCR5, AKT, p-AKT, and TNF-α and the co-expression of IBA1 and CCR5 in the corpus callosum of the mice were detected with Western blotting and immunofluorescence double staining. In a BV2 microglial cell model of oxygen-glucose deprivation (OGD), the effects of pretreatment with gastrodin and Maraviroc (an CCR5 antagonist) on protein expressions of CCR5, AKT, p-AKT, TNF-α and IL-1β were evaluated using Western blotting and immunofluorescence double staining.
Results:
The neonatal mice with HIBD showed significantly increased expressions of CCR5 and TNF-α with lowered p-AKT expression in the brain tissues, and GAS treatment obviously reversed these changes. HIBD also significantly increased the co-expression of IBA1 and CCR5 in the corpus callosum of the mice, which was obviously lowered by gastrodin treatment. In BV2 cells, OGD significantly increased the expressions of CCR5, TNF-α, and IL-1β and decreased the expression of p-AKT, and these changes were inhibited by treatment with gastrodin, Maraviroc or their combination; the inhibitory effect of the combined treatment did not differ significantly from that of gastrodin or Maraviroc alone.
Conclusion:
Gastrodin can produce neuroprotective effects in neonatal mice with HIBD by inhibiting inflammatory cytokine production and activate AKT phosphorylation via inhibiting CCR5.
Insights
Gastrodin protects neonatal mouse brains from hypoxic-ischemic brain damage (HIBD) by reducing inflammation via CCR5 inhibition. This mechanism involves activating AKT phosphorylation, offering neuroprotection.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Context:
- Hypoxic-ischemic brain damage (HIBD) is a significant cause of neonatal neurological deficits.
- Microglia-mediated inflammation plays a crucial role in HIBD pathogenesis.
- Gastrodin, a traditional Chinese medicine component, has shown potential neuroprotective properties.
Purpose:
- To elucidate the mechanism by which gastrodin exerts neuroprotection against HIBD in neonatal mice.
- To investigate the role of CCR5 and AKT signaling in gastrodin's anti-inflammatory effects.
- To evaluate gastrodin's impact on microglia activation and inflammatory cytokine production.
Summary:
- Neonatal mice subjected to HIBD exhibited increased CCR5 and TNF-α expression and decreased p-AKT levels.
- Gastrodin treatment reversed these molecular changes in vivo and in vitro (BV2 microglial cells).
- Gastrodin, along with the CCR5 antagonist Maraviroc, inhibited OGD-induced inflammatory responses in BV2 cells, suggesting CCR5 as a key target.
Impact:
- This study reveals gastrodin's neuroprotective mechanism in HIBD through CCR5 inhibition and AKT activation.
- Findings suggest gastrodin as a potential therapeutic agent for neonatal HIBD.
- Understanding the role of CCR5 in HIBD opens new avenues for targeted therapeutic interventions.
