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Defining a Time Window for Neuroprotection and Glia Modulation by Caffeine After Neonatal Hypoxia-Ischaemia
Elena Di Martino1, Erica Bocchetta2,3, Shunichiro Tsuji2,4
1Department of Women's and Children's Health, Karolinska Institutet, BioClinicum J9:30 Visionsgatan 4, 17176, Stockholm, Sweden. elena.di.martino@ki.se.
Insights
Early caffeine administration immediately after hypoxic-ischemic (HI) brain injury in neonatal mice reduced brain damage and inflammation. Prompt treatment normalized behavioral deficits, highlighting the critical timing for caffeine
Area of Science:
- Neonatal neurology
- Neuroscience
- Pharmacology
Background:
- Hypoxic-ischemic (HI) brain injury is a major cause of neonatal brain damage with lasting effects.
- Caffeine, an adenosine receptor inhibitor, treats preterm apnea.
Purpose of the Study:
- To investigate the neuroprotective effects of caffeine administered at different times after HI in neonatal mice.
- To evaluate caffeine's impact on brain morphology, inflammation, and behavior post-HI.
Main Methods:
- Neonatal mouse pups (P10) received caffeine at 0, 6, 12, or 24 hours post-HI.
- Behavioral tests (open field, rotarod) and brain morphology were assessed 2 weeks post-injury.
- Gene expression and immunohistology (MAP2, MBP, microglia, apoptosis, astrogliosis) were analyzed at 1 and 5 days post-HI.
Main Results:
- A single caffeine dose immediately after HI significantly reduced grey and white matter lesions.
- Caffeine decreased microglia, apoptotic cells, astrogliosis, and pro-inflammatory cytokine expression.
- Immediate caffeine administration normalized hyperactivity observed in HI mice; later doses showed no benefit.
Conclusions:
- Caffeine demonstrates neuroprotection and immunomodulation in neonatal HI brain injury only when given immediately after the insult.
- Timing of intervention is critical for caffeine's efficacy in this neonatal brain injury model.
Abstract:
Hypoxic-ischemic (HI) brain injury remains an important cause of brain damage in neonates with potential life-long consequences. Caffeine, which is a competitive inhibitor of adenosine receptors, is commonly used as treatment for preterm apnoea in clinical settings. In the current study, we investigated the effects of caffeine given at 0 h, 6 h, 12 h or 24 h after HI in P10 mouse pups. Open field and rotarod behavioural tests were performed 2 weeks after injury, and brain morphology was then evaluated. Gene expression and immunohistological analyses were assessed in mice 1- and 5-day post-HI. A single dose of caffeine directly after HI resulted in a reduction of the lesion in the grey and white matter, judged by immunostaining of MAP2 and MBP, respectively, compared to PBS-treated controls. In addition, the number of amoeboid microglia and apoptotic cells, the area covered by astrogliosis, and the expression of pro-inflammatory cytokines were significantly decreased. Behavioural assessment after 2 weeks showed increased open-field activity after HI, and this was normalised if caffeine was administered immediately after the injury. Later administrations of caffeine did not change the outcomes when compared to the vehicle group. In conclusion, caffeine only yielded neuroprotection and immunomodulation in a neonatal model of brain hypoxia ischaemia if administered immediately after injury.
