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.

中文翻译：

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定义新生儿缺氧缺血后咖啡因对神经保护和神经胶质细胞调节的时间窗。

缺氧缺血性 (HI) 脑损伤仍然是新生儿脑损伤的一个重要原因，并具有潜在的终生后果。咖啡因是腺苷受体的竞争性抑制剂，在临床环境中常用于治疗早产呼吸暂停。在目前的研究中，我们研究了 P10 小鼠幼崽在 HI 后 0 小时、6 小时、12 小时或 24 小时给予咖啡因的影响。损伤后 2 周进行旷场和旋转杆行为测试，然后评估脑形态。HI 后 1 天和 5 天在小鼠中评估基因表达和免疫组织学分析。与 PBS 处理的对照相比，HI 后直接单剂量的咖啡因导致灰质和白质中的病变减少，分别通过 MAP2 和 MBP 的免疫染色来判断。此外，变形小胶质细胞和凋亡细胞的数量、星形胶质细胞增生覆盖的面积以及促炎细胞因子的表达均显着降低。2 周后的行为评估显示 HI 后开放场活动增加，如果在受伤后立即服用咖啡因，这会正常化。与载体组相比，后来服用咖啡因并没有改变结果。总之，如果在受伤后立即给药，咖啡因仅在新生儿脑缺氧缺血模型中产生神经保护和免疫调节作用。如果在受伤后立即服用咖啡因，这就会正常化。与载体组相比，后来服用咖啡因并没有改变结果。总之，如果在受伤后立即给药，咖啡因仅在新生儿脑缺氧缺血模型中产生神经保护和免疫调节作用。如果在受伤后立即服用咖啡因，这就会正常化。与载体组相比，后来服用咖啡因并没有改变结果。总之，如果在受伤后立即给药，咖啡因仅在新生儿脑缺氧缺血模型中产生神经保护和免疫调节作用。