METHODS: To identify genes associated with cerebral I/R injury, high-throughput sequencing was employed. The potential downstream genes were further analyzed using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Protein-Protein Interaction (PPI) analyses. For experimental models, primary microglia and neurons were extracted from the cortical tissues of mouse brains. An in vitro cerebral I/R injury model was established in microglia using the oxygen-glucose deprivation/reoxygenation (OGD/R) technique. In vivo models involved inducing cerebral I/R injury in mice through the middle cerebral artery occlusion (MCAO) method. These models were used to assess neurological function, immune cell infiltration, and inflammatory factor release.
RESULTS: The study identified RPS27A as a key player in cerebral I/R injury, with PSMD12 likely acting as its downstream regulator. Silencing RPS27A in OGD/R-induced microglia decreased the release of inflammatory factors and reduced neuron apoptosis. Additionally, RPS27A silencing in cerebral cortex tissues mediated the PSMD12/NF-κB axis, resulting in decreased inflammatory factor release, reduced neutrophil infiltration, and improved cerebral injury outcomes in I/R-injured mice.
CONCLUSIONS: RPS27A regulates the expression of the PSMD12/NF-κB signaling axis, leading to the induction of inflammatory factors in microglial cells, promoting immune cell infiltration in brain tissue, and exacerbating brain damage in I/R mice. This study introduces novel insights and theoretical foundations for the treatment of nerve damage caused by I/R, suggesting that targeting the RPS27A and downstream PSMD12/NF-κB signaling axis for drug development could represent a new direction in I/R therapy.
方法:为了鉴定与脑I/R损伤相关的基因,采用高通量测序。使用基因本体论(GO)进一步分析了潜在的下游基因,京都基因和基因组百科全书(KEGG),和蛋白质-蛋白质相互作用(PPI)分析。对于实验模型,从小鼠大脑皮层组织中提取原代小胶质细胞和神经元。使用氧糖剥夺/复氧(OGD/R)技术在小胶质细胞中建立了体外脑I/R损伤模型。体内模型涉及通过大脑中动脉闭塞(MCAO)方法在小鼠中诱导脑I/R损伤。这些模型用于评估神经功能,免疫细胞浸润,和炎症因子释放。
结果:研究确定RPS27A是大脑I/R损伤的关键角色,PSMD12可能充当其下游监管机构。在OGD/R诱导的小胶质细胞中沉默RPS27A减少了炎症因子的释放并减少了神经元凋亡。此外,大脑皮层组织RPS27A沉默介导PSMD12/NF-κB轴,导致炎症因子释放减少,中性粒细胞浸润减少,并改善I/R损伤小鼠的脑损伤结果。
结论:RPS27A调节PSMD12/NF-κB信号轴的表达,导致小胶质细胞炎症因子的诱导,促进脑组织中的免疫细胞浸润,并加剧I/R小鼠的脑损伤。这项研究为I/R引起的神经损伤的治疗提供了新的见解和理论基础。提示靶向RPS27A和下游PSMD12/NF-κB信号轴进行药物开发可能代表I/R治疗的新方向。