Sepsis encephalopathy (SAE) has a high incidence and mortality rate in patients with sepsis; however, there is currently no effective treatment. Our previous studies have reported that 2% hydrogen (H₂) gas inhalation had a protective effect on sepsis and SAE; however, the specific mechanism have not been fully elucidated. In the current study, male Institute of Cancer Research mice were either used to create the cecal ligation and puncture (CLP) model or for sham surgery, followed by 2% H₂ gas inhalation for 60 min beginning at 1 and 6 h following sham or CLP surgeries. The isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, hematoxylin and eosin (H&E) staining, Nissl staining, and western blot analysis were used to investigate the effects of H₂ on brain injury in mice with sepsis. The results of the H&E, and Nissl staining indicated that the CLP mice had a significant brain injury, which was characterized by aggravated pathological damage and was alleviated by 2% H₂ inhalation. Quantitative proteomics based on iTRAQ combined with LC-MS/MS analysis quantified a total of 5317 proteins, of which 39 were connected with the protective mechanism of H₂. In addition, H₂ could regulate the immune and the coagulation systems. Furthermore, western blot analysis revealed that H₂ decreased SAE in septic mice by downregulating the protein expression levels of SMAD4, DPYS, PTGDS and upregulating the expression level of CUL4A. These results provide insights into the mechanism of the positive effect of H₂ on SAE and contribute to the clinical application of H₂ in patients with sepsis.

脓毒症脑病（SAE）在脓毒症患者中的发病率和死亡率都很高；但是，目前没有有效的治疗方法。我们之前的研究报道，2% 的氢气 (H ₂ ) 气体吸入对败血症和 SAE 有保护作用；但具体机制尚未完全阐明。在目前的研究中，雄性癌症研究所小鼠要么用于创建盲肠结扎和穿刺 (CLP) 模型，要么用于假手术，然后使用 2% H ₂在假手术或 CLP 手术后 1 和 6 小时开始吸入气体 60 分钟。基于相对和绝对定量 (iTRAQ) 的定量蛋白质组学的同量异位标记结合液相色谱-串联质谱 (LC-MS/MS) 分析、苏木精和伊红 (H&E) 染色、Nissl 染色和蛋白质印迹分析用于研究 H ₂对脓毒症小鼠脑损伤的影响。H&E和Nissl染色结果表明CLP小鼠有明显的脑损伤，其特点是病理损伤加重，吸入2%H ₂后减轻。基于iTRAQ的定量蛋白质组学结合LC-MS/MS分析共定量了5317种蛋白质，其中39种与H的保护机制有关。₂ . 此外，H ₂可以调节免疫和凝血系统。此外，蛋白质印迹分析显示，H ₂通过下调 SMAD4、DPYS、PTGDS 的蛋白表达水平和上调 CUL4A 的表达水平来降低脓毒症小鼠的 SAE。这些结果提供了对 H ₂对 SAE积极作用机制的见解，并有助于 H ₂在败血症患者中的临床应用。