Downregulation of<italic style="font-style: italic"> PFN1 </italic>exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress <italic style="font-style: italic">via</italic> regulating <italic style="font-style: italic">RhoA/ROCK2 </italic>signaling pathway

JIANG Ting; ZHENG Qian; DUAN Kui; CHEN Changling; PENG Shuang; LIU Ying; WANG Jifen; ZHANG Chunlin; YE Lan; DAI Zhijun; FENG Zhanhui

doi:10.16016/j.2097-0927.202512010

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Downregulation of PFN1 exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress via regulating RhoA/ROCK2 signaling pathway

Monographic Reports | 更新时间：2026-05-30

- Downregulation of PFN1 exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress via regulating RhoA/ROCK2 signaling pathway
- Journal of Army Medical University Vol. 48, Issue 10, Pages: 1298-1312(2026)
- 作者机构：
  
  1.贵州医科大学附属医院神经内科，贵州贵阳
  2.贵州医科大学基础医学院，药理学教研室贵州贵阳
  3.贵州医科大学基础医学院，生物教研室，贵州贵阳
  4.贵州省人民医院神经内科，贵州贵阳
- 作者简介：
  
  FENG Zhanhui， E-mail：h9450203@126.com
- 基金信息：
  
  the Regional Science Program of National Natural Science Foundation of China(81960224;82360266);the Project of Guizhou Provincial Department of Science and Technology Plan Project;the PhD Research Startup Fund of the Affiliated Hospital of Guizhou Medical University(gyfybsky-2025-33)
- DOI：10.16016/j.2097-0927.202512010
  CLC： R341;R394.3;R742.1
- Received：02 December 2025，
  
  Revised：2026-01-10，
  
  Accepted：16 January 2026，
  
  Published：30 May 2026
- 稿件说明：
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江婷，郑乾，段逵，等.下调PFN1通过调控RhoA/ROCK2信号通路稳定钙稳态并抑制氧化应激发挥癫痫神经保护作用［J］.陆军军医大学学报，2026，48（10）：1298-1312.DOI：10.16016/j.2097-0927.202512010.

JIANG T，ZHENG Q，DUAN K，et al.Downregulation of PFN1 exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress via regulating RhoA/ROCK2 signaling pathway［J］.J Army Med Univ，2026，48（10）：1298-1312.DOI：10.16016/j.2097-0927.202512010.
江婷，郑乾，段逵，等.下调PFN1通过调控RhoA/ROCK2信号通路稳定钙稳态并抑制氧化应激发挥癫痫神经保护作用［J］.陆军军医大学学报，2026，48（10）：1298-1312.DOI：10.16016/j.2097-0927.202512010. DOI：

JIANG T，ZHENG Q，DUAN K，et al.Downregulation of PFN1 exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress via regulating RhoA/ROCK2 signaling pathway［J］.J Army Med Univ，2026，48（10）：1298-1312.DOI：10.16016/j.2097-0927.202512010. DOI：

摘要

目的

癫痫是全球最常见的神经系统疾病之一，其发病机制涉及神经元网络异常与氧化损伤，目前缺乏有效干预策略，探索新靶点具有重要意义。本研究旨在探究下调肌动蛋白结合蛋白1（profilin 1，PFN1）通过调控

RhoA

ROCK2

信号通路稳定癫痫海马神经元内钙稳态和抑制神经元的氧化应激。

方法

通过基因表达综合（Gene Expression Omnibus，GEO）数据库和人类蛋白质图谱（the Human Protein Atlas，THPA）数据库，开展生物信息学分析，以探究颞叶癫痫的关键差异表达基因。体内实验中，将82只雄性8~10周龄C57BL/6J小鼠（体质量20~25 g）按随机数字表法分为对照组、癫痫模型组、空载对照组及

PFN1

干扰组。其中，空载对照组与

PFN1

干扰组分别经脑立体定向注射siCon或si

PFN1

复合物，72 h后注射红藻氨酸（kainic acid，KA）诱导癫痫模型；其余2组仅给予生理盐水和KA。进一步采用RT-qPCR及Western blotting实验验证海马神经元

PFN1

表达特征；通过脑电图和Racine评分评估

PFN1

下调后癫痫模型鼠癫痫发作特征，通过旷场实验评估小鼠自发运动能力和探索行为水平；采用RT-qPCR、Western blotting、免疫组化染色及组织染色等方法分析

PFN1

下调后对海马神经元损伤的保护效应。体外实验中，提前对HT22细胞进行

PFN1

干扰，按相同分组采用无镁细胞外液建立癫痫细胞模型。通过钙成像、活性氧（reactive oxygen species，ROS）探针、流式细胞术等方法评估钙稳态和氧化应激水平，并通过免疫共沉淀和Western blotting实验验证

PFN1

与

RhoA/ROCK2

信号通路的相互作用。

结果

通过生物信息学分析，发现在癫痫模型中

PFN1

表达显著上调；表征结果显示，在癫痫模型中，

PFN1

转录及蛋白表达均显著升高（

0.01），且与NeuN共定位于海马CA1、CA3区神经元细胞质内。敲低

PFN1

可显著降低Racine评分（

0.001）、延长发作潜伏期（

0.001），改善癫痫样放电及神经元形态，增加NeuN阳性细胞及尼氏小体数量（

0.01）。机制研究表明，PFN1与RhoA、ROCK2形成蛋白复合物，敲低

PFN1

可抑制该通路蛋白表达（

0.05），下调CAMK2水平（

0.05），显著抑制钙内流（

0.001）及ROS生成（

0.001）。

结论

下调

PFN1

可能通过

RhoA

ROCK2

信号通路稳定癫痫海马神经元内钙稳态和抑制神经元氧化应激从而达到保护神经元效应。

Abstract

Objective

Epilepsy is one of the most common neurological disorders worldwide

with pathogenesis involving abnormal neuronal networks and oxidative damage. Effective interventions remain limited

highlighting the importance of exploring novel targets. This study aimed to investigate how downregulation of profilin-1 (PFN1) exerts neuroprotective effects by stabilizing calcium homeostasis and suppressing oxid

ative stress in hippocampal neurons

via

the

RhoA/ROCK2

signaling pathway in epilepsy.

Methods

Bioinformatics analysis was performed using the Gene Expression Omnibus (GEO) and the Human Protein Atlas (THPA) databases to identify key differentially expressed genes in temporal lobe epilepsy. For in vivo experiments

82 male C57BL/6 mice (aged 8 to10 weeks

weighting 20 to 25 g) were divided into control

epileptic model

vector control

and

PFN1

-interfered groups. The vector control and

PFN1-

interfered groups received stereotactic injections of siCon or

siPFN1

complexes

respectively

followed by kainic acid (KA)-induced epilepsy modeling 72 h later; the other 2 groups received saline and KA only. RT-qPCR and Western blotting validated

PFN1

expression in hippocampal neurons.Electroencephalogram and Racine scores were used to evaluate seizure characteristics and electroencephalographic features. Open field tests assessed spontaneous locomotion and exploratory behavior. RT-qPCR

Western blotting

immunohistochemistry

and histological staining analyzed

PFN1

’s protective effects against hippocampal neuronal injury. For

in vitro

experiments

HT22 cells were pretreated with

PFN1

interference before establishing the epileptic cell models using Mg

-free extracellular fluid under identical conditions

with calcium imaging

reactive oxygen species (ROS) probes

and flow cytometry assessing calcium homeostasis and oxidative stress. Co-immunoprecipitation and Western blotting verified

PFN1

’s interaction with the

RhoA/ROCK2

pathway.

Results

Bioinformatic analyses revealed significant upregulation of

PFN1

expression in epileptic models. Results from characterization demonstrated elevated

PFN1

transcription and protein expression (

0.01) in epileptic model

with

PFN1

co-localized with NeuN in the cytoplasm of hippocampal CA1/CA3 neurons.

PFN1

knockdown significantly reduced Racine scores (

0.001)

prolonged seizure latency (

0.001)

improved epileptiform discharges and neuronal morphology

and increased NeuN-positive cells and Nissl bodies (

0.01). Mechanistically

PFN1

formed a complex with RhoA and ROCK2; Its knockdown suppressed pathway protein expression (

0.05)

downregulated CaMK2 levels (

0.05)

and inhibited calcium influx (

0.001) and ROS generation (

0.001). Conclusion Downregulation of

PFN1

may protect neurons in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress through the

RhoA/ROCK2

signaling pathway.

关键词

Keywords

references

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Related Institution

Department of Occupational Health， School of Public Health， Shanxi Medical University

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⁰

Downregulation of PFN1 exerts neuroprotective effects in epilepsy by stabilizing calcium homeostasis and suppressing oxidative stress via regulating RhoA/ROCK2 signaling pathway

DOI：10.16016/j.2097-0927.202512010

摘要

Abstract

关键词

Keywords

references