Olfactory mucosal mesenchymal stem cells inhibit ferroptosis and attenuate cerebral ischemia-reperfusion injury by up-regulating GPX4

ZHUANG Junhong; YANG Guoshuai; PENG Jun; CHEN Zigui; TANG Hong; XIA Ying

doi:10.16016/j.2097-0927.202503064

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Olfactory mucosal mesenchymal stem cells inhibit ferroptosis and attenuate cerebral ischemia-reperfusion injury by up-regulating GPX4

Neuroscience | 更新时间：2025-07-14

- Olfactory mucosal mesenchymal stem cells inhibit ferroptosis and attenuate cerebral ischemia-reperfusion injury by up-regulating GPX4
- Journal of Army Medical University Vol. 47, Issue 13, Pages: 1420-1428(2025)
- 作者机构：
  
  1.中南大学湘雅医学院附属海口医院：，神经内科，海南海口
  2.中南大学湘雅医学院附属海口医院：，神经外科，海南海口
- 作者简介：
  
  XIA Ying， E-mail： xiaying008@163.com
- 基金信息：
  
  the Regional Science Fund Program of National Natural Science Foundation of China(82460268);the Joint Project of Health Science and Technology Innovation of Hainan Province(WSJK2024MS186);the Special Fund of Hainan Provincial Clinical Research Center for Cerebrovascular Diseases(LCYX202309)
- DOI：10.16016/j.2097-0927.202503064
  CLC： R338.1;R345;R743.31
- Received：31 December 2024，
  
  Revised：03 April 2025，
  
  Published：15 July 2025
- 稿件说明：
移动端阅览
庄俊鸿，杨国帅，彭俊，等.嗅觉黏膜间充质干细胞通过上调GPX4抑制铁死亡减轻脑缺血再灌注损伤［J］.陆军军医大学学报，2025，47（13）：1420-1428.DOI：10.16016/j.2097-0927.202503064.

ZHUANG J H，YANG G S，PENG J，et al.Olfactory mucosal mesenchymal stem cells inhibit ferroptosis and attenuate cerebral ischemia-reperfusion injury by up-regulating GPX4［J］.J Army Med Univ，2025，47（13）：1420-1428.DOI：10.16016/j.2097-0927.202503064.
庄俊鸿，杨国帅，彭俊，等.嗅觉黏膜间充质干细胞通过上调GPX4抑制铁死亡减轻脑缺血再灌注损伤［J］.陆军军医大学学报，2025，47（13）：1420-1428.DOI：10.16016/j.2097-0927.202503064. DOI：

ZHUANG J H，YANG G S，PENG J，et al.Olfactory mucosal mesenchymal stem cells inhibit ferroptosis and attenuate cerebral ischemia-reperfusion injury by up-regulating GPX4［J］.J Army Med Univ，2025，47（13）：1420-1428.DOI：10.16016/j.2097-0927.202503064. DOI：

摘要

目的

探究嗅觉黏膜间充质干细胞（olfactory mucosa mesenchymal stem cells，OM-MSCs）通过谷胱甘肽氧化酶4（glutathione peroxidase 4，GPX4）改善氧糖剥夺/复氧（oxygen and glucose deprivation/reperfusion，OGD/R）诱导的神经元细胞铁死亡。

方法

收集鼻息肉患者的中鼻道正常嗅觉黏膜组织，并提取OM-MSCs，采用流式细胞仪检测人嗅觉黏膜分离培养的原代细胞表面标志物CD34、CD45、CD73、CD90、CD105、CD146，显微镜下观察细胞形态学。将小鼠海马神经元细胞HT22细胞按随机数字表法分为：对照组、OGD/R组、OGD/R+OM-MSCs组、OGD/R+sh-NC组、OGD/R+sh-GPX4组、OGD/R+sh-GPX4+OM-MSCs组。对HT22细胞进行OGD/R处理，并给予OM-MSCs共培养和/或GPX4敲低处理。采用流式细胞仪检测HT22细胞凋亡，CCK-8法检测HT22细胞活力，试剂盒检测HT22细胞中铁死亡相关指标MDA、R

OS、GSH、Fe

，qPCR及Western blot检测GPX4表达。

结果

人嗅觉黏膜分离培养的原代细胞表面CD34及CD45呈阴性，CD73、CD90、CD105、CD146呈阳性，细胞贴壁生长并呈梭状，细胞表面标志物及形态学符合OM-MSCs特征。在HT22细胞方面，对照组、OGD/R组、OGD/R+OM-MSCs组在细胞活力、MDA、ROS、GSH、Fe

、GPX4方面具有统计学差异（

0.05）；OGD/R组与对照组相比，细胞活力和GSH降低（

0.05），MDA、ROS、Fe

升高（

0.05），GPX4下调（

0.05）；OGD/R+OM-MSCs组与OGD/R组相比（

0.05），细胞活力和GSH升高（

0.05），MDA、ROS、Fe

降低（

0.05），GPX4上调。OGD/R+sh-GPX4组与OGD/R-sh-NC组相比，细胞活力和GSH降低（

0.05），MDA、ROS、Fe

升高（

0.05），并伴随有GPX4下调（

0.05），OM-MSCs可逆转以上变化。

结论

OM-MSCs通过上调GPX4在HT22细胞中的表达抑制OGD/R诱导的HT22细胞铁死亡。

Abstract

Objective‍　‍To investigate whether olfactory mucosa mesenchymal stem cells (OM-MSCs) attenuate oxygen glucose deprivation and recovery (OGD/R)-induced ferroptosis in neurons through glutathione oxidase 4 (GPX4). Methods‍　‍The middle nasal tissue were collected from a patient with nasal polyps admitted in our hospital

and then OM-MSCs were isolated from the tissue

which were confirmed by morphological observation under light microscopy and phenotypic characterization through flow cytometry for surface markers

including CD34

CD45

CD73

CD90

CD105

and CD146. Mouse hippocampal neuronal cell line HT22 was randomly divided into control

Control

OGD/R

OGD/R+OM-MSCs

OGD/R+sh-NC

OGD/R+sh-GPX4 and OGD/R+sh-GPX4+OM-MSCs groups. After the cells were subjected to OGD/R modeling

the cells were subsequently co-cultured with OM-MSCs and/or knockdown of GPX4. Neuronal apoptosis was quantified by flow cytometry

while cell viability was assessed using CCK-8 assay. Biochemical markers associated with ferroptosis

including MDA

ROS

GSH

and Fe²⁺ levels

were measured with corresponding reagent kits. The GPX4 expression at

both mRNA and protein levels was determined through qPCR and Western blotting

respectively. Results‍　‍The isolated and primarily cultured OM-MSCs showed typical characteristics of OM-MSCs in cell surface markers (negative expression of CD34 and CD45 but positive expression of CD73

CD90

CD105

and CD146 on cell surface) and morphology (adherent cells in a spindle-like shape). Significant differences were observed among the control

OGD/R

and OGD/R+OM-MSCs groups in terms of cell viability

MDA

ROS

GSH

and GPX4 (

0.05). The OGD/R group showed notable decreases in cell activity and GSH (

0.05)

increases in MDA

ROS

and Fe

(

0.05)

and down-regulation of GPX4 when compared with the control group (

005). Co-culture with OM-MSCs enhanced cell activity and GSH (

0.05)

decreased MDA

ROS

and Fe

(

0.05)

and up-regulated GPX4 as compared to the conditions in the OGD/R group (

0.05). While

OGD/R+sh-GPX4 treament developed the decreases in cell viability

GSH

and GPX4 and the increases in MDA

ROS

and Fe

as compared to the OGD/R+sh-NC group (

0.05)

however

all of these could be reversed by OM-MSCs. Conclusion‍　‍OM-MSCs inhibit OGD/R-induced ferroptosis in HT22 cells by up-regulating GPX4.

关键词

Keywords

references

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