RNA-seq of rats in the Sham, ICH (Intracerebral hemorrhage), and EDB (Edaravone dexborneol) groups

Edaravone dexborneol (EDB) is widely recognized for its anti-inflammatory and antioxidant properties and is clinically applied in the treatment of acute cerebral infarction. Ferroptosis is a critical process in the pathophysiology of brain injury following intracerebral hemorrhage (ICH). However, it remains unclear whether EDB can ameliorate ICH through the modulation of ferroptosis. This study aimed to evaluate the function and mechanism of EDB in treatment of ICH. With a rat ICH model, animal behavior tests, histopathological staining, magnetic resonance imaging and evans blue staining were used to evaluate the neural protective function of EDB on ICH rats. The potential molecular mechanism was investigated using RNA sequencing. With the administration of Fer-1, a range of ferroptosis-related biomarkers, including Fe2+, 4-hydroxynonenal, malondialdehyde, etc., were analyzed to to ascertain whether EDB confers neuroprotective effects through the modulation of P53/GPX4 pathways to inhibit ferroptosis. Finally, the findings were further corroborated using an in vitro ICH model with a P53 inhibitor. EDB has the potential to markedly enhance nerve and motor function, mitigate pathological damage, facilitate hematoma clearance, and repair BBB injury in ICH rats. KEGG analysis revealed that the differentially expressed genes were associated with signaling pathways, including P53 and ferroptosis. Both EDB and Fer-1 substantially reduced the concentrations of Fe2+, 4-hydroxynonenal, malondialdehyde, increased the amount of anti-oxidants, decreased the expression of P53, and concurrently upregulated the expression of GPX4. Besides, the P53 inhibitor PFT-α was observed to significantly reduce the levels of 4-HNE and lipid peroxides, while concurrently increasing the expression of GPX4. This investigation has shed light on the crucial neuroprotective role of EDB by regulating ferroptosis in ICH disease, which provided a theoretical basis for the clinical application of EDB in the treatment of ICH.依达拉奉右旋莰醇（Edaravone dexborneol, EDB）因兼具抗炎与抗氧化活性而广受认可，目前已被临床应用于急性脑梗死的治疗。铁死亡（Ferroptosis）是脑出血（intracerebral hemorrhage, ICH）后脑损伤病理生理过程中的关键环节，但目前尚不清楚EDB是否可通过调控铁死亡改善脑出血损伤。本研究旨在探讨EDB治疗脑出血的作用及其分子机制。本研究通过构建大鼠脑出血模型，采用动物行为学实验、组织病理染色、磁共振成像以及伊文思蓝（Evans Blue）染色等手段，评估EDB对脑出血大鼠的神经保护作用；通过转录组测序（RNA sequencing）探究其潜在分子机制；通过给予铁死亡抑制剂Fer-1，检测一系列铁死亡相关生物标志物（包括Fe²+、4-羟基壬烯醛（4-hydroxynonenal）、丙二醛（malondialdehyde）等）的水平，以明确EDB是否通过调控P53/GPX4通路抑制铁死亡，从而发挥神经保护作用。最后，本研究通过构建体外脑出血模型并联合P53抑制剂，进一步验证了上述实验结果。实验结果表明，EDB可显著改善脑出血大鼠的神经与运动功能，减轻病理损伤，促进血肿清除，并修复血脑屏障（Blood-Brain Barrier, BBB）损伤。京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes, KEGG）富集分析显示，差异表达基因主要富集于P53通路与铁死亡通路等信号通路。EDB与Fer-1均可显著降低Fe²+、4-羟基壬烯醛、丙二醛的水平，提升抗氧化物质含量，下调P53的表达，同时上调谷胱甘肽过氧化物酶4（GPX4）的表达。此外，P53抑制剂PFT-α可显著降低4-羟基壬烯醛与脂质过氧化物的水平，同时上调GPX4的表达。本研究揭示了EDB通过调控铁死亡在脑出血疾病中发挥关键神经保护作用的机制，为EDB临床应用于脑出血治疗提供了坚实的理论依据。