GenX induces cochlear hair cell injury <italic style="font-style: italic">via</italic> System Xc⁻-Gpx4-Fsp1 ferroptosis axis

LIANG Zhengwei; HONG Huihui; YUAN Wei

doi:10.16016/j.2097-0927.202507094

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GenX induces cochlear hair cell injury via System Xc⁻-Gpx4-Fsp1 ferroptosis axis

Basic Medicine | 更新时间：2025-11-13

- GenX induces cochlear hair cell injury via System Xc⁻-Gpx4-Fsp1 ferroptosis axis
- Journal of Army Medical University Vol. 47, Issue 21, Pages: 2652-2662(2025)
- 作者机构：
  
  1.西南医科大学附属医院耳鼻咽喉头颈外科，四川泸州
  2.重庆市人民医院（重庆大学附属人民医院）耳鼻咽喉头颈外科，重庆
  3.重庆大学医学院神经智能中心实验室，重庆
- 作者简介：
  
  YUAN Wei， E-mail： weiyuan175@sina.com
- 基金信息：
  
  the General Program of National Natural Science Foundation of China(81873702);the Open Research Project of National Clinical Research Center for Otorhinolaryngology(2024KF008);the Key Special Program for Technological Innovation and Application Development of Chongqing(CSTB2023TIAD-KPX0059);the Major Project of Chongqing Health Commission(2022DBXM006)
- DOI：10.16016/j.2097-0927.202507094
  CLC： R334.16;R363.21;R994.6
- Received：28 July 2025，
  
  Revised：2025-10-23，
  
  Accepted：23 October 2025，
  
  Published：15 November 2025
- 稿件说明：
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梁正薇，洪慧慧，袁伟.GenX通过System Xc⁻-Gpx4-Fsp1铁死亡轴诱导耳蜗毛细胞损伤［J］.陆军军医大学学报，2025，47（21）：2652-2662.DOI：10.16016/j.2097-0927.202507094.

LIANG Z W，HONG H H，YUAN W.GenX induces cochlear hair cell injury via System Xc⁻-Gpx4-Fsp1 ferroptosis axis［J］.J Army Med Univ，2025，47（21）：2652-2662.DOI：10.16016/j.2097-0927.202507094.
梁正薇，洪慧慧，袁伟.GenX通过System Xc⁻-Gpx4-Fsp1铁死亡轴诱导耳蜗毛细胞损伤［J］.陆军军医大学学报，2025，47（21）：2652-2662.DOI：10.16016/j.2097-0927.202507094. DOI：

LIANG Z W，HONG H H，YUAN W.GenX induces cochlear hair cell injury via System Xc⁻-Gpx4-Fsp1 ferroptosis axis［J］.J Army Med Univ，2025，47（21）：2652-2662.DOI：10.16016/j.2097-0927.202507094. DOI：

摘要

目的

探讨铁死亡在2，3，3，3-四氟-2-（七氟丙氧基）丙酸铵［ammonium 2，3，3，3-tetrafluoro-2-（heptafluoropropoxy）-propanoate，GenX］暴露诱导耳蜗毛细胞损伤中的作用与机制。

方法

体外培养HEI-OC1细胞，分为对照组、GenX组、铁死亡抑制剂（ferrostatin-1，Fer-1）组、GenX+Fer-1组，CCK-8检测GenX对耳蜗毛细胞的毒性作用，CCK-8检测GenX+Fer-1共处理后观察有无铁死亡的发生，Western blot和qRT-PCR检测铁死亡标志物、耳蜗分化功能指标

、

耳血迷路屏障标志物和听力相关铁死亡通路蛋白和基因的表达，使用FerroOrange、Bodipy（C11）、线粒体膜电位荧光探针（mitochondrial membrane potential assay kit，JC-1）和ATP探针分别检测Fe²⁺蓄积、脂质过氧化水平、线粒体膜损伤和ATP情况。

结果

200 μmol/L GenX暴露12 h后显著降低HEI-OC1细胞活力（

0.01）。GenX暴露12 h后引起磷脂氢过氧化物谷胱甘肽过氧化物酶（phospholipid hydroperoxide glutathione peroxidase，Gpx4）、溶质载体家族7成员11和铁死亡抑制蛋白1（ferroptosis suppressor protein 1，Fsp1）蛋白表达下调（

0.05），酰基辅酶A合成酶长链家族成员4蛋白表达上调（

0.01），耳蜗毛细胞分化功能基因和耳血迷路屏障基因表达降低（

0.05）

，

同时伴随Fe²⁺蓄积、脂质过氧化水平升高和线粒体膜损伤及降低ATP生成（

0.001）。GenX+Fer-1组可显著恢复细胞活力（

0.05），逆转铁死亡蛋白的表达（

0.05），并改善部分耳蜗分化功能基因和耳血迷路屏障基因的表达（

0.05）。同时GenX+Fer-1组可减少Fe²⁺蓄

积、降低脂质过氧化水平、抑制线粒体膜损伤和升高ATP水平（

0.001）。

结论

GenX通过诱导铁代谢紊乱、脂质过氧化蓄积，并依赖System Xc⁻-Gpx4-Fsp1铁死亡轴途径损伤耳蜗毛细胞的分化和屏障功能。

Abstract

Objective

To investigate the role and mechanism of ferroptosis in cochlear hair cell injury induced by exposure to ammonium 2

3-tetrafluoro-2-(heptafluoropropoxy)propanoate (GenX).

Methods

Mouse cochlear hair cell line HEI-OC1 was assigned to control

GenX

ferrostatin-1 (Fer-1)

and GenX+Fer-1 groups. CCK-8 assay was used to assess the cytotoxicity of GenX and the rescue effect of Fer-1 co-treatment. Western blotting and qRT-PCR were employed to measure the protein and transcriptional expression of ferroptosis markers

cochlear function indicators

blood labyrinth barrier markers

and ferroptosis-related pathway. FerroOrange

Bodipy(C11)

mitochondrial membrane potential assay kit (JC-1)

and adenosine triphosphate (ATP) assay were applied to detect Fe²⁺ accumulation

lipid peroxidation

mitochondrial membrane dysfunction

and cellular ATP levels

respectively.

Results

Exposure to 200 μmol/L GenX for 12 h significantly reduced the viability of HEI-OC1 cells (

0.01)

down-regulated the protein levels of glutathione peroxidase 4 (Gpx4)

solute carrier family 7 member 11 (Slc7a11)

and ferroptosis suppressor protein 1 (Fsp1) (

0.05)

whereas up-regulated acyl-coa synthetase long-chain family member 4 (Acsl4) (

0.01)

and decreased the expression of cochlear hair-cell function genes and blood labyrinth barrier genes (

0.05). These changes were accompanied with Fe

accumulation

elevated lipid peroxidation

mitochondrial membrane damage

and reduced ATP production (

0.001). Addition of Fer-1 restored cell viability (

0.05)

restored the expression of ferroptosis

related proteins (

0.05)

and improved the expression of several hair-cell function and blood labyrinth barrier genes (

0.05). In parallel

the GenX+Fer-1 group exhibited reduced Fe²⁺ accumulation

lower lipid peroxidation

attenuated mitochondrial membrane damage

and increased ATP level (

0.001).

Conclusion

GenX induces iron-metabolism dysregulation and lipid peroxidation

and then leads to differentiation impairment of cochlear hair cells and barrier functions

via

the ferroptotic System Xc⁻-Gpx4-Fsp1 axis.

关键词

Keywords

references

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GenX induces cochlear hair cell injury via System Xc⁻-Gpx4-Fsp1 ferroptosis axis

DOI：10.16016/j.2097-0927.202507094

摘要

Abstract

关键词

Keywords

references