Liu Chang, Li Zhixi, Xi Hongjie
Heilongjiang Province Key Laboratory of Research on Anesthesiology and Critical Care Medicine, Harbin, China.
The Key Laboratory of Myocardial Ischemia Organization, Chinese Ministry of Education, Harbin, China.
Front Pharmacol. 2022 Nov 24;13:940260. doi: 10.3389/fphar.2022.940260. eCollection 2022.
Ischemic stroke (IS) is a neurological condition associated with high mortality and disability rates. Although the molecular mechanisms underlying IS remain unclear, ferroptosis was shown to play an important role in its pathogenesis. Hence, we applied bioinformatics analysis to identify ferroptosis-related therapeutic targets in IS. IS-related microarray data from the GSE61616 dataset were downloaded from the Gene Expression Omnibus (GEO) database and intersected with the FerrDb database. In total, 33 differentially expressed genes (DEGs) were obtained and subjected to functional enrichment and protein-protein interaction (PPI) network analyses. Four candidate genes enriched in the HIF-1 signaling pathway (, , , and ) were selected based on the hierarchical clustering of the PPI dataset. We also downloaded the IR-related GSE35338 dataset and GSE58294 dataset from the GEO database to verify the expression levels of these four genes. ROC monofactor analysis demonstrated a good performance of , , , and in the diagnosis of ischemic stroke. Transcriptional levels of the above four genes, and translational level of GPX4, the central regulator of ferroptosis, were verified in a mouse model of middle cerebral artery occlusion (MCAO)-induced IS by qRT-PCR and western blotting. Considering the regulation of the HIF-1 signaling pathway, dexmedetomidine was applied to the MCAO mice. We found that expression of these four genes and GPX4 in MCAO mice were significantly reduced, while dexmedetomidine reversed these changes. In addition, dexmedetomidine significantly reduced MCAO-induced cell death, improved neurobehavioral deficits, and reduced the serum and brain levels of inflammatory factors (TNF-α and IL-6) and oxidative stress mediators (MDA and GSSG). Further, we constructed an mRNA-miRNA-lncRNA network based on the four candidate genes and predicted possible transcription factors. In conclusion, we identified four ferroptosis-related candidate genes in IS and proposed, for the first time, a possible mechanism for dexmedetomidine-mediated inhibition of ferroptosis during IS. These findings may help design novel therapeutic strategies for the treatment of IS.
缺血性中风（IS）是一种与高死亡率和致残率相关的神经系统疾病。尽管IS潜在的分子机制尚不清楚，但铁死亡在其发病机制中起着重要作用。因此，我们应用生物信息学分析来识别IS中与铁死亡相关的治疗靶点。从基因表达综合数据库（GEO）下载了来自GSE61616数据集的IS相关微阵列数据，并与FerrDb数据库进行交叉分析。总共获得了33个差异表达基因（DEG），并对其进行了功能富集和蛋白质-蛋白质相互作用（PPI）网络分析。基于PPI数据集的层次聚类，选择了四个富集在HIF-1信号通路中的候选基因（、、和）。我们还从GEO数据库下载了IR相关的GSE35338数据集和GSE58294数据集，以验证这四个基因的表达水平。ROC单因素分析表明，、、和在缺血性中风诊断中表现良好。通过qRT-PCR和蛋白质印迹法在大脑中动脉闭塞（MCAO）诱导的IS小鼠模型中验证了上述四个基因的转录水平以及铁死亡的核心调节因子GPX4的翻译水平。考虑到HIF-1信号通路的调节作用，将右美托咪定应用于MCAO小鼠。我们发现MCAO小鼠中这四个基因和GPX4的表达显著降低，而右美托咪定逆转了这些变化。此外，右美托咪定显著减少了MCAO诱导的细胞死亡，改善了神经行为缺陷，并降低了血清和大脑中炎症因子（TNF-α和IL-6）以及氧化应激介质（MDA和GSSG）的水平。此外，我们基于这四个候选基因构建了一个mRNA-miRNA-lncRNA网络，并预测了可能的转录因子。总之，我们在IS中鉴定出四个与铁死亡相关的候选基因，并首次提出了右美托咪定介导的IS期间铁死亡抑制的可能机制。这些发现可能有助于设计治疗IS的新策略。
