METHODS: The SAH models were constructed by injection of arterial blood into cisterna Magana. Three groups of C57 mice were randomly assigned: Sham, SAH, and BHB. All mice were subjected to neurological function assessment, western blot, immunofluorescence double staining, and RNA-seq. Glymphatic system function was examined with tracer and immunofluorescence double staining, and the differential genes were examined with RNA-seq. In addition, the expression of related inflammation was detected.
RESULTS: Compared with the SAH group, BHB reinstated AQP4 polarization by upregulating SNTA1 protein to enhance the glymphatic system. According to RNA-seq, the different genes were primarily connected to microglia activation, astrocytes, and inflammation. Western blot and immunofluorescence further confirmed that the related inflammatory protein expression levels were upregulated. BHB attenuated neuroinflammation after SAH. Ultimately, it can mitigate the neurological deficits in SAH mice.
CONCLUSIONS: The study reveals a novel mechanism that BHB treatment mitigates neurologic impairment in SAH mice. We propose that BHB may play a neuroprotective effect by enhancing glymphatic system function and attenuating neuroinflammatory subarachnoid hemorrhage.
方法:SAH模型通过将动脉血注射到大羊角池构建。三组C57小鼠被随机分配:假,SAH,还有BHB.对所有小鼠进行神经功能评估,westernblot,免疫荧光双重染色,和RNA-seq。用示踪剂和免疫荧光双重染色检查淋巴系统功能,用RNA-seq检测差异基因。此外,检测相关炎症的表达。
结果:与SAH组相比,BHB通过上调SNTA1蛋白以增强淋巴系统来恢复AQP4极化。根据RNA-seq,不同的基因主要与小胶质细胞的激活有关,星形胶质细胞,和炎症。Westernblot和免疫荧光进一步证实相关炎性蛋白表达水平上调。BHB减轻SAH后的神经炎症。最终,它可以减轻SAH小鼠的神经功能缺损。
结论:该研究揭示了BHB治疗减轻SAH小鼠神经损伤的新机制。我们认为BHB可能通过增强淋巴系统功能和减轻神经炎性蛛网膜下腔出血来发挥神经保护作用。