High-Throughput mRNA Sequencing Reveals Potential Therapeutic Targets of Febuxostat in Secondary Injury After Intracerebral Hemorrhage. High-Throughput mRNA Sequencing Reveals Potential Therapeutic Targets of Febuxostat in Secondary Injury After Intracerebral Hemorrhage

Febuxostat is a urate-lowering medication for the treatment of patients with gout. This study was performed to elucidate the effects and underlying mechanisms of febuxostat on neuronal injury induced by intracerebral hemorrhage (ICH) in mice. The results showed that the administration of febuxostat improved neurological severity scores and blood-brain barrier (BBB) permeability. Moreover, febuxostat attenuated neuronal cell death and cytokine levels compared with the ICH group. Next, we conducted a transcriptome analysis of the neuroprotective effects of febuxostat. The overlapping significant differentially expressed genes (DEGs) were identified. Gene ontology (GO) analysis revealed that the overlapping significant DEGs were most enriched in five items. The intersecting DEGs of the aforementioned five pathways were Wisp1, Wnt7b, Frzb, and Pitx2. In addition, GO terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways revealed that DEGs were mainly involved in the wnt signaling pathway. Furthermore, the expression of Wisp1 and Wnt7b in the perihematomal region at 72 h post-ICH was observed. The results showed that both Wisp1 and Wnt7b were increased in the ICH group and were decreased by the administration of febuxostat. Taken together, the study showed that febuxostat protected against secondary brain injury after ICH and the Wnt7b-Wisp1 pathway was closely related to neuroprotective effects. Overall design: Differentially expressed genes were identified among Sham, ICH and Feb groups by using mRNA-Seq

非布司他（Febuxostat）是一种用于治疗痛风患者的降尿酸药物。本研究旨在阐明非布司他对小鼠脑出血（intracerebral hemorrhage, ICH）诱导的神经元损伤的作用及其潜在机制。结果显示，给予非布司他可改善神经功能严重程度评分，并降低血脑屏障（blood-brain barrier, BBB）通透性。相较于ICH模型组，非布司他可减轻神经元细胞死亡，同时降低细胞因子水平。随后，我们针对非布司他的神经保护作用开展了转录组分析，鉴定得到了重叠的显著差异表达基因（differentially expressed genes, DEGs）。基因本体论（Gene ontology, GO）分析显示，这些重叠的显著DEGs在5个条目上富集程度最高。上述5条通路的交集差异表达基因为Wisp1、Wnt7b、Frzb及Pitx2。此外，GO条目及京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）通路分析表明，差异表达基因主要参与Wnt信号通路。我们进一步检测了ICH造模后72小时血肿周围区域Wisp1与Wnt7b的表达情况，结果显示ICH模型组中Wisp1与Wnt7b的表达均上调，而非布司他给药可降低二者的表达水平。综上，本研究证实非布司他可对抗脑出血后的继发性脑损伤，且Wnt7b-Wisp1通路与其神经保护作用密切相关。实验整体设计：采用信使RNA测序（mRNA-Seq）技术，对假手术组（Sham）、ICH模型组及非布司他给药组（Feb组）的样本进行差异表达基因筛选。