Mechanism of transcription factor SP1 regulating ferroptosis in early brain injury after subarachnoid hemorrhage

TANG Qi-kai; ZHOU Xiao-ming; ZHANG Bing-tao; MA Chen-feng; WU Qi; CHEN Shu-juan; WU Wei; ZHANG Xin

doi:10.11659/jjssx.04E025132

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Mechanism of transcription factor SP1 regulating ferroptosis in early brain injury after subarachnoid hemorrhage

Basic Research | 更新时间：2025-12-22

- Mechanism of transcription factor SP1 regulating ferroptosis in early brain injury after subarachnoid hemorrhage
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 34, Issue 12, Pages: 1069-1076(2025)
- 作者机构：
  
  1.南京大学医学院附属金陵医院神经外科，江苏南京 210002
  2.南京医科大学第一附属医院神经外科，江苏南京 210029
- 作者简介：
- 基金信息：
- DOI：10.11659/jjssx.04E025132
  CLC： R743.35
- Received：24 April 2025，
  
  Published：25 December 2025
- 稿件说明：
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汤其凯,周晓明,张冰涛等.转录因子SP1调控蛛网膜下腔出血后早期脑损伤铁死亡的机制研究[J].局解手术学杂志,2025,34(12):1069-1076.

TANG Qi-kai,ZHOU Xiao-ming,ZHANG Bing-tao,et al.Mechanism of transcription factor SP1 regulating ferroptosis in early brain injury after subarachnoid hemorrhage[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(12):1069-1076.
汤其凯,周晓明,张冰涛等.转录因子SP1调控蛛网膜下腔出血后早期脑损伤铁死亡的机制研究[J].局解手术学杂志,2025,34(12):1069-1076. DOI： 10.11659/jjssx.04E025132.

TANG Qi-kai,ZHOU Xiao-ming,ZHANG Bing-tao,et al.Mechanism of transcription factor SP1 regulating ferroptosis in early brain injury after subarachnoid hemorrhage[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(12):1069-1076. DOI： 10.11659/jjssx.04E025132.

摘要

目的

探讨转录因子SP1在蛛网膜下腔出血（SAH）后早期脑损伤中的表达及其对神经元铁死亡的调控作用。

方法

构建小鼠SAH模型及体外原代神经元OxyHb刺激模型，通过Western blot、qPCR、免疫组化染色及免疫荧光染色检测SP1的表达变化。通过体外敲低和过表达SP1，检测SP1蛋白表达、细胞活性、乳酸脱氢酶（LDH）水平及丙二醛（MDA）、4-羟基壬烯醛（4-HNE）、谷胱甘肽（GSH）、Fe²⁺铁死亡相关指标，并在不同细胞死亡抑制剂处理后检测敲低SP1的细胞活性变化。在体内过表达SP1后，通过尼氏染色、脑水肿程度、改良Garcia评分、平衡木实验、水迷宫实验评估其对神经功能的保护作用。通过双荧光素酶报告基因实验及分子检测验证SP1对GPX4表达的转录调控作用。

结果

在SAH后早期，SP1的表达显著下调。在体外敲低SP1后，神经元中MDA、4-HNE及Fe²⁺水平升高（

0.05），GSH水平下降（

0.05），而过表达SP1则逆转了上述变化。铁死亡抑制剂干预能够提高敲低SP1后的细胞活性，而其他抑制剂处理后细胞活性无显著变化。体内过表达SP1后，小鼠脑水肿程度改善，神经认知功能恢复。SP1通过结合GPX4启动子区域正向调控其表达，发挥抑制神经元铁死亡作用。

结论

在SAH早期，过表达SP1可转录激活GPX4抑制神经元铁死亡，具有显著的神经保护作用，为SAH后的干预治疗提供了潜在分子靶点。

Abstract

Objective

To investigate the expression of transcription factor SP1 in early brain injury after subarachnoid hemorrhage （SAH） and its regulatory effect on neuronal ferroptosis.

Methods

A mouse model of SAH and an in vitro primary neuronal model with OxyHb stimulation were constructed. The change of SP1 expression was detected by Western blot，qPCR，immunohistochemistry，and immunofluorescence staining. The expression of SP1 protein，cell activity，lactate dehydrogenase （LDH） level，and ferroptosis-related indicators malondialdehyde （MDA），4-hydroxynonenal （4-HNE），glutathione （GSH） and Fe²⁺ were detected after knockdown and overexpression of SP1 in vitro，and the viability changes of cells with SP1 knockdown were detected after treated with different cell death inhibitors. The neuroprotective effects of SP1 after its overexpression in vivo were assessed through Nissl staining，the degree of brain edema，modified Garcia score，balance beam test，and water maze test. The transcriptional regulatory effect of SP1 on GPX4 expression was verified through dual luciferase reporter gene experiment and molecular detection.

Results

The expression of SP1 was significantly downregulated during early brain injury after SAH. After knockdown of SP1 in vitro，the levels of MDA，4-HNE and Fe²⁺ in neurons increased （

0.05），GSH levels decreased （

0.05），while overexpression of SP1 in vitro reversed these changes. The activity of cells with SP1 knockdown was increased after intervention with ferroptosis inhibitors，whereas the cell viability showed no significant change after treated with other inhibitors. Overexpression of SP1 in vivo ameliorated brain edema，and recovered neurological cognitive function. SP1 positively regulated the expression of GPX4 by binding to its promoter region，thereby exerting an inhibitory effect on neuronal ferroptosis.

Conclusion

In the early stage of SAH，overexpression of SP1 can transcriptionally activate GPX4 to inhibit neuronal ferroptosis，which has a significant neuroprotective effect and provides a potential molecular target for intervention and treatment after SAH.

关键词

Keywords

references

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