Hu Qianxin, Zeng Haixin, Feng Chengao, Tian Wei, He Yuxin, Li Bing
Department of Neurosurgery, Jiangnan University Medical Center, Wuxi Neurosurgical Institute, 68 Zhongshan Road, Wuxi, Jiangsu Province, 214002, P.R. China.
Exp Anim. 2025 Jan 10;74(1):39-48. doi: 10.1538/expanim.24-0078. Epub 2024 Aug 7.
Secondary brain injury (SBI) is one of the main causes of high mortality and disability rates following intracerebral hemorrhage (ICH). Tumor necrosis factor receptor-associated factor 6 (TRAF6) plays a crucial role in the process of pyroptosis, and modulating its expression may present a novel therapeutic strategy for mitigating brain injury. This study aims to explore the mechanisms of TRAF6 in pyroptosis after ICH. C57BL/6J mice were used to establish the ICH model. Brain was collected at different time points for q-PCR and western blot to detect the level of TRAF6. After the C25-140 (the TRAF6 inhibitor) was administrated, the mice were divided into four groups. Then, the neurological deficit, brain water content, and blood-brain barrier (BBB) damage were detected. Immunofluorescence and western blot were used to detect the level of pyroptosis proteins, and ELISA and q-PCR were used to detect the levels of IL-18 and IL-1β. TRAF6 expression was upregulated after ICH and was mainly expressed in neurons. Inhibition of TRAF6 expression with C25-140 alleviated neurological deficits and reduced brain edema after ICH. In addition, inhibition of TRAF6 also reduced the expression of pyroptosis inflammasomes such as GSDMD, NLRP3, and ASC, as well as neurological damage caused by IL-18 and IL-1β after ICH. TRAF6 regulates neuronal pyroptosis in SBI after ICH. Inhibition of TRAF6 may be a potential target for alleviating inflammatory damage after ICH.
继发性脑损伤(SBI)是脑出血(ICH)后高死亡率和高致残率的主要原因之一。肿瘤坏死因子受体相关因子6(TRAF6)在细胞焦亡过程中起关键作用,调节其表达可能为减轻脑损伤提供一种新的治疗策略。本研究旨在探讨TRAF6在脑出血后细胞焦亡中的作用机制。采用C57BL/6J小鼠建立脑出血模型。在不同时间点采集脑组织进行q-PCR和蛋白质免疫印迹检测TRAF6水平。给予C25-140(TRAF6抑制剂)后,将小鼠分为四组。然后,检测神经功能缺损、脑含水量和血脑屏障(BBB)损伤情况。采用免疫荧光和蛋白质免疫印迹检测细胞焦亡蛋白水平,采用酶联免疫吸附测定(ELISA)和q-PCR检测白细胞介素-18(IL-18)和白细胞介素-1β(IL-1β)水平。脑出血后TRAF6表达上调,主要表达于神经元。用C25-140抑制TRAF6表达可减轻脑出血后的神经功能缺损并减轻脑水肿。此外,抑制TRAF6还可降低细胞焦亡炎性小体如gasdermin D(GSDMD)、NLR家族含pyrin结构域蛋白3(NLRP3)和凋亡相关斑点样蛋白(ASC)的表达,以及脑出血后IL-18和IL-1β引起的神经损伤。TRAF6调节脑出血后SBI中的神经元细胞焦亡。抑制TRAF6可能是减轻脑出血后炎症损伤的潜在靶点。