Department of Neurology, The First Affiliated Hospital of Harbin Medical University, Harbin, China.
Sci Rep. 2024 Sep 19;14(1):21860. doi: 10.1038/s41598-024-67118-2.
No single treatment significantly reduces the mortality rate and improves neurological outcomes after intracerebral haemorrhage (ICH). New evidence suggests that pyroptosis-specific proteins are highly expressed in the perihaematomal tissues of patients with ICH and that the disulfiram (DSF) inhibits pyroptosis. An ICH model was established in C57BL/6 mice by intracranial injection of collagenase, after which DSF was used to treat the mice. Cell model of ICH was constructed, and DSF was used to treat the cells. HE, TUNEL, Nissl, FJC and IF staining were performed to evaluate the morphology of brain tissues; Western blotting and ELISA were performed to measure the protein expression of NOD-like receptor protein 3 (NLRP3)/Caspase-1/gasdermin D (GSDMD) classical pyroptosis pathway and Toll-likereceptor4 (TLR4)/nuclear factor-kappaB (NF-κB) inflammatory signaling pathway and blood‒brain barrier-associated factoes, and the wet/dry weight method was used to determine the brain water content. The expression of proteins related to the NLRP3/Caspase-1/GSDMD pathway and the TLR4/NF-κB pathway was upregulated in tissues surrounding the haematoma compared with that in control tissues; Moreover, the expression of the blood-brain barrier structural proteins occludin and zonula occludens-1 (ZO-1) was downregulated, and the expression of Aquaporin Protein-4 (AQP4) and matrix metalloprotein 9 (MMP-9) was upregulated. DSF significantly inhibited these changes, reduced the haematoma volume, decreased the brain water content, reduced neuronal death and degeneration and improved neurological function after ICH. ICH activated the classical pyroptosis pathway and TLR4/NF-κB inflammatory pathway, disruped the expression of blood-brain barrier structural proteins, and exacerbated brain injury and neurological dysfunction. DSF inhibited these changes and exerted the therapeutic effects on pathological changes and dysfunction caused by ICH.
没有单一的治疗方法能显著降低脑出血（ICH）患者的死亡率和改善神经功能结局。新证据表明，细胞焦亡特异性蛋白在脑出血患者血肿周围组织中高度表达，而双硫仑（DSF）抑制细胞焦亡。采用胶原酶脑内注射法建立 C57BL/6 小鼠 ICH 模型，然后用 DSF 治疗小鼠。构建 ICH 细胞模型，用 DSF 处理细胞。行 HE、TUNEL、尼氏染色、FJC 和免疫荧光染色观察脑组织形态；Western blot 和 ELISA 法检测 NOD 样受体蛋白 3（NLRP3）/半胱氨酸天冬氨酸蛋白酶-1（Caspase-1）/Gasdermin D（GSDMD）经典细胞焦亡通路和 Toll 样受体 4（TLR4）/核因子-κB（NF-κB）炎症信号通路及血脑屏障相关因子蛋白表达，干湿重法测定脑水含量。与对照组相比，血肿周围组织中 NLRP3/Caspase-1/GSDMD 通路和 TLR4/NF-κB 通路相关蛋白表达上调；血脑屏障结构蛋白紧密连接蛋白（occludin）和闭合蛋白-1（ZO-1）表达下调，水通道蛋白 4（AQP4）和基质金属蛋白酶 9（MMP-9）表达上调。DSF 可显著抑制这些改变，减少血肿体积，降低脑水含量，减少神经元死亡和变性，改善 ICH 后神经功能。ICH 激活经典细胞焦亡通路和 TLR4/NF-κB 炎症通路，破坏血脑屏障结构蛋白表达，加重脑损伤和神经功能障碍。DSF 抑制这些改变，对 ICH 引起的病理变化和功能障碍发挥治疗作用。
