Numerous studies have shown that local excessive inflammatory response in brain tissue was an important pathogenesis of secondary injury following cerebral ischemia-reperfusion (I/R). However, the inflammatory-related targets and pathways after cerebral I/R injury are still unclear. This study was to investigate possible targets and mechanisms after cerebral I/R injury. Rats were subjected to transient or permanent middle cerebral artery occlusion (MCAO). Neurological deficit scores test was used to evaluate neurological function. Cerebral infarction was evaluated by MRI, TTC staining and Nissl staining. Microglia activation was detected by immunofluorescence using Iba-1 antibody. Inflammatory factors were detected by ELISA assay. RNA-sequencing transcriptome analysis was processed and the differential genes were verified by real-time quantitative PCR (qPCR) and western blotting. The results showed that neurological function of rats in I/R group was more severe than that in I group on the 7th after cerebral I/R. Therefore, the differences between cerebral ischemia and cerebral I/R for 7 days were studied in further study. The results showed that the levels of pro-inflammatory factors in I/R group were higher and the levels of anti-inflammatory factors were lower than those in I group. KEGG pathway and gene network enrichment analysis revealed that some common differential up- and down-regulated genes were involved in most of significant pathways. These common differential up-regulated genes belonged to TLR4/MYD88 inflammatory signaling pathway and common differential down-regulated genes belonged to HRAS/RAF1 neurotrophic signaling pathway. Interestingly, according to the genetic interaction analysis of string database, these up-regulated differential genes might promote the development of inflammation, while the down-regulated differential genes might inhibit the development of inflammation. Furthermore, qPCR and WB results verified that these pro-inflammatory genes in the I/R group were higher than those in the I group, while possible anti-inflammatory genes in the I/R group were lower than those in the I group. It is concluded that TLR4/MYD88 inflammatory signaling pathway and HRAS/RAF1 neurotrophic signaling pathway may play different roles after cerebral I or I/R and may be therapeutic targets for stroke recovery.

中文翻译：

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脑缺血再灌注加重大鼠脑梗死损伤及RNA-Seq鉴定的可能差异基因。

大量研究表明，脑组织局部过度炎症反应是脑缺血再灌注（I/R）继发性损伤的重要发病机制。然而，脑 I/R 损伤后炎症相关的靶点和通路尚不清楚。本研究旨在探讨脑 I/R 损伤后可能的靶点和机制。大鼠经受短暂或永久性大脑中动脉闭塞 (MCAO)。神经功能缺损评分测试用于评估神经功能。通过MRI、TTC染色和Nissl染色评估脑梗塞。使用 Iba-1 抗体通过免疫荧光检测小胶质细胞活化。ELISA法检测炎症因子。处理 RNA 测序转录组分析，并通过实时定量 PCR (qPCR) 和蛋白质印迹验证差异基因。结果显示，脑I/R后第7天，I/R组大鼠的神经功能较I组严重。因此，在进一步研究中研究了7天脑缺血和脑I/R之间的差异。结果显示，I/R组促炎因子水平高于I组，抗炎因子水平低于I组。KEGG通路和基因网络富集分析表明，一些常见的差异上调和下调基因参与了大多数重要通路。这些常见的差异上调基因属于TLR4/MYD88炎症信号通路，常见的差异下调基因属于HRAS/RAF1神经营养信号通路。有趣的是，根据字符串数据库的遗传相互作用分析，这些上调的差异基因可能促进炎症的发展，而下调的差异基因可能抑制炎症的发展。此外，qPCR和WB结果证实I/R组中这些促炎基因高于I组，而I/R组中可能的抗炎基因低于I组。