Ferrostatin-1下调脑缺血再灌注小鼠NF-κB/MMP-9维持AQP4极性分布减轻脑水肿

doi:10.3969/j.issn.1006-7795.2026.02.012

首都医科大学学报 ›› 2026, Vol. 47 ›› Issue (2): 308-315.doi: 10.3969/j.issn.1006-7795.2026.02.012

• 脑血管病的基础与临床研究 • 上一篇下一篇

Ferrostatin-1下调脑缺血再灌注小鼠NF-κB/MMP-9维持AQP4极性分布减轻脑水肿

师文娟,郭正然,陈小东,胡妍姝,戚智锋^*

首都医科大学宣武医院北京市老年病医疗研究中心脑心病学研究室,北京 100053

收稿日期:2025-12-16 修回日期:2026-01-13 出版日期:2026-04-21 发布日期:2026-04-21
通讯作者: 戚智锋 E-mail:qizhifeng@xwh.ccmu.edu.cn
基金资助:
国家自然科学基金项目（82271308），北京市自然科学基金项目（7222080）。

Ferrostatin-1 attenuates cerebral edema by down-regulating NF-κB/MMP-9 and preserving AQP4 polarization in mice after ischemia-reperfusion

Shi Wenjuan, Guo Zhengran, Chen Xiaodong, Hu Yanshu, Qi Zhifeng^*

Research Laboratory of Neuro Cardio Vascular Diseases, Beijing Geriatric Medical Research Center, Xuanwu Hospital, Capital Medical University, Beijing 100053, China

Received:2025-12-16 Revised:2026-01-13 Online:2026-04-21 Published:2026-04-21
Supported by:
This study was supported by National Natural Science Foundation of China (82271308)， Natural Science Foundation of Beijing(7222080).

摘要/Abstract

摘要： 目的研究铁死亡抑制剂ferrostatin-1 (Fer-1)对大脑中动脉闭塞（middle cerebral artery occlusion, MCAO）再灌注小鼠脑组织核因子-κB（nuclear factor kappa B, NF-κB）、基质金属蛋白酶-9（matrix metalloproteinase-9, MMP-9）及水通道蛋白-4（aquaporin-4, AQP4）的影响，探究抑制铁死亡对缺血性脑卒中后水肿的作用及其可能的机制。方法将健康成年雄性C57BL/6J小鼠采用数字表法随机分为 3 组（每组n=10~13）：假手术组（Sham组）、MCAO + vehicle组（MCAO组）、MCAO + Fer-1组（Fer-1组）。使用改良线栓法制作MCAO模型，缺血60 min后拔栓实现再灌注。于再灌注即刻，Fer-1组尾静脉注射Fer-1（0.8 mg/kg），Sham组和MCAO组尾静脉注射等体积溶剂。各组动物于再灌注后24 h处死取材，测量梗死侧脑组织干湿质量计算脑含水量；Western blotting 检测小鼠缺血脑组织中NF-κB、MMP-9及AQP4的表达；免疫荧光双标染色检测AQP4在CD31标记的血管周围的极化表达。使用Image J软件进行Western blotting 条带的灰度分析及AQP4荧光染色的极化分析。结果 ①与 Sham 组相比，MCAO 组小鼠梗死侧脑组织含水量显著增加（P＜0.01）；Fer-1组缺血脑组织含水量较MCAO 组明显降低（P＜0.05）。②MCAO 组小鼠缺血侧脑组织 NF-κB、MMP-9及AQP4总蛋白水平比 Sham 组明显升高（P＜0.05）；与 MCAO 组相比，Fer-1组缺血侧脑组织 NF-κB、MMP-9蛋白水平显著降低（P＜0.01），AQP4总蛋白表达明显降低（P＜0.05）。③与 Sham 组相比，MCAO 组小鼠缺血脑组织AQP4 M23/M1比值显著下降（P＜0.01）；Fer-1组比值较MCAO组明显升高（P＜0.05）。④MCAO 组小鼠缺血侧血管处AQP4信号峰值与背景信号比值显著低于 Sham 组（P＜0.01）；给予Fer-1的缺血小鼠AQP4信号峰值与背景信号比值较MCAO 组显著升高（P＜0.01）。结论铁死亡抑制剂 Fer-1可以下调NF-κB、MMP-9和AQP4的表达水平，维持AQP4极性分布，减轻小鼠脑缺血再灌注24 h的脑水肿。

关键词: ferrostatin-1, 脑缺血, 脑水肿, 核因子-κB（NF-κB）, 基质金属蛋白酶-9（MMP-9）, 水通道蛋白-4（AQP4）极化

Abstract: Objective To investigate whether the ferroptosis inhibitor ferrostatin-1 (Fer-1) attenuates cerebral edema 24 h after middle-cerebral-artery occlusion-reperfusion (MCAO/R) in mice and to explore the underlying mechanisms focusing on nuclear factor kappa B (NF-κB), matrix metalloproteinase-9 (MMP-9) and aquaporin-4 (AQP4) polarity. Methods Adult male C57BL/6J mice were randomly allocated to Sham, MCAO/R + vehicle, and MCAO/R + Fer-1 groups (n=10－13 per group). Sixty-minute filament occlusion was followed by reperfusion. Fer-1 (0.8 mg·kg-1) or equal-volume vehicle was administered intravenously at the onset of reperfusion. Twenty-four hours later, brain water content was quantified by the dry-wet weight method; Western blotting was employed to detect total NF-κB, MMP-9 and AQP4 levels as well as the AQP4-M23/M1 isoform ratio; and immunofluorescence co-labelling of AQP4 with CD31 was performed to assess perivascular AQP4 polarization. Image J was used for densitometric analysis of Western blotting bands and quantitative assessment of fluorescence intensity. Results ① Brain water content was markedly elevated in the MCAO/R group compared with Sham (P<0.01), and this increase was significantly attenuated by Fer-1 (P<0.05). ②The protein expression levels of NF-κB, MMP-9 and total AQP4 were up-regulated in the ischemic hemisphere after MCAO/R (P<0.05 versus Sham); Fer-1 treatment substantially decreased NF-κB and MMP-9 expression (P<0.01) and moderately down-regulated total AQP4 (P<0.05). ③The AQP4-M23/M1 ratio, an index of AQP4 polarization, was significantly decreased after MCAO/R (P<0.01 versus Sham) and was restored by Fer-1 (P<0.05).④ In the ischemic hemisphere, the peak-to-background fluorescence ratio of perivascular AQP4 was lower in MCAO/R mice than that in Sham mice (P<0.05); Fer-1 administration reversed this deficit (P<0.05). Conclusion Fer-1 attenuates early cerebral edema following MCAO/R by suppressing NF-κB and MMP-9 signaling, down-regulating total AQP4 abundance and preserving perivascular AQP4 polarization. These findings highlight ferroptosis inhibition as a potential therapeutic strategy for ischemia－reperfusion-induced brain edema.

Key words: ferrostatin-1, cerebral ischemia, brain edema, nuclear factor kappa B（NF-κB）, matrix metalloproteinase-9（MMP-9）, aquaporin-4（AQP4）polarization

中图分类号:

R743.3

师文娟, 郭正然, 陈小东, 胡妍姝, 戚智锋. Ferrostatin-1下调脑缺血再灌注小鼠NF-κB/MMP-9维持AQP4极性分布减轻脑水肿[J]. 首都医科大学学报, 2026, 47(2): 308-315.

Shi Wenjuan, Guo Zhengran, Chen Xiaodong, Hu Yanshu, Qi Zhifeng. Ferrostatin-1 attenuates cerebral edema by down-regulating NF-κB/MMP-9 and preserving AQP4 polarization in mice after ischemia-reperfusion[J]. Journal of Capital Medical University, 2026, 47(2): 308-315.

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Ferrostatin-1下调脑缺血再灌注小鼠NF-κB/MMP-9维持AQP4极性分布减轻脑水肿

Ferrostatin-1 attenuates cerebral edema by down-regulating NF-κB/MMP-9 and preserving AQP4 polarization in mice after ischemia-reperfusion

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