Zhao Gengshui, Zhao Jianfei, Lang Jiadong, Sun Guozhu
Department of Neurosurgery, Harrison International Peace Hospital Affiliated to Hebei Medical University, Hengshui, China.
Department of Neurosurgery, The People's Hospital of Shijiazhuang City, Shijiazhuang, China.
Mol Neurobiol. 2025 Feb;62(2):1894-1903. doi: 10.1007/s12035-024-04371-6. Epub 2024 Jul 24.
Pyroptosis has been implicated in many pathologic processes, including endoplasmic reticulum stress (ERS). However, the underlying mechanisms and molecular targets of ERS affecting pyroptosis still need further exploration. We obtained gene sets associated with ERS and pyroptosis, and the common genes were regarded as crosstalk genes linking ERS and pyroptosis. Protein-protein interaction (PPI) network was constructed, and the hub genes were obtained via Cytoscape. Moreover, to validate the efficacy of the therapeutic target, neurological tests, brain water content measurements, Nissl staining, Western blot, ELISA, TUNEL analyses, and transmission electron microscopy were performed in a mouse model. A total of 13 crosstalk genes were acquired, and enrichment analysis revealed that these genes were mainly enriched in stress-associated cellular processes and pathways, including KEAP1-NFE2L2 pathway. The hub gene, NFE2L2, was identified by Cytoscape, and tert-butylhydroquinone (tBHQ) was screened as candidate drug to activate NFE2L2. Western blot and ELISA results showed that activation of NFE2L2 could attenuate the expression of ERS and pyroptosis-related proteins by promoting nuclear translocation of Nrf2 (encoded by NFE2L2). Pathological evaluation by Nissl staining and TUNEL assay reflected a similar trend. Furthermore, activation of NFE2L2 ameliorated neurological deficits and reduced brain edema. In conclusion, our bioinformatic analysis results established the theoretical foundation of NFE2L2 as a promising therapeutic target. Moreover, in the mouse model, tBHQ pretreatment further confirmed the effectiveness of this target. We hypothesized NFE2L2 may play a key role in the progression of ERS-mediated pyroptosis. These findings may inspire new ideas to treat neurological disorders.
细胞焦亡与包括内质网应激(ERS)在内的许多病理过程有关。然而,ERS影响细胞焦亡的潜在机制和分子靶点仍需进一步探索。我们获得了与ERS和细胞焦亡相关的基因集,将共同基因视为连接ERS和细胞焦亡的串扰基因。构建了蛋白质-蛋白质相互作用(PPI)网络,并通过Cytoscape获得了枢纽基因。此外,为了验证治疗靶点的有效性,在小鼠模型中进行了神经学测试、脑含水量测量、尼氏染色、蛋白质印迹、酶联免疫吸附测定、TUNEL分析和透射电子显微镜检查。共获得13个串扰基因,富集分析表明这些基因主要富集于与应激相关的细胞过程和途径,包括KEAP1-NFE2L2途径。通过Cytoscape鉴定出枢纽基因NFE2L2,并筛选出叔丁基对苯二酚(tBHQ)作为激活NFE2L2的候选药物。蛋白质印迹和酶联免疫吸附测定结果表明,激活NFE2L2可通过促进Nrf2(由NFE2L2编码)的核转位来减弱ERS和细胞焦亡相关蛋白的表达。尼氏染色和TUNEL分析的病理评估反映了类似的趋势。此外,激活NFE2L2改善了神经功能缺损并减轻了脑水肿。总之,我们的生物信息学分析结果奠定了NFE2L2作为一个有前景的治疗靶点的理论基础。此外,在小鼠模型中,tBHQ预处理进一步证实了该靶点的有效性。我们推测NFE2L2可能在ERS介导的细胞焦亡进程中起关键作用。这些发现可能为治疗神经疾病激发新的思路。