解读缺血性中风中的铁死亡：关键基因与针灸的治疗潜力

Decoding ferroptosis in ischemic stroke: key genes and the therapeutic potential of acupuncture.

作者信息

Wu Chunxiao, Xu Zhirui, Wang Qizhang, Zhu Shuping, Tang Chunzhi

机构信息

Shenzhen Hospital of Integrated Traditional Chinese and Western Medicine, Shenzhen, China.

Shenzhen Clinical College of Integrated Chinese and Western Medicine, Guangzhou University of Chinese Medicine, Shenzhen, Guangdong, China.

出版信息

Front Aging Neurosci. 2025 Jun 9;17:1506276. doi: 10.3389/fnagi.2025.1506276. eCollection 2025.

DOI:10.3389/fnagi.2025.1506276

PMID:40552324

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC12183209/

Abstract

BACKGROUND

Ferroptosis has been reported to be associated with the development of ischemic stroke; however, comprehensive investigations of ferroptosis-associated genes are lacking. Herein, key differentially expressed genes (DEGs) related to ferroptosis in ischemic stroke were identified and validated and the potential mechanism of acupuncture in treating ischemic stroke was explored through bioinformatics analysis and experiments.

METHODS

High-throughput RNA sequencing was performed to identify DEGs between cerebral ischemic injury tissue and normal brain tissue in mice. Subsequently, differentially expressed ferroptosis-related genes (DE-FRGs) were identified by intersecting the DEGs with a ferroptosis database. Functional enrichment analysis was conducted to investigate potential signaling pathways involved in ischemic stroke, while protein-protein interaction (PPI) analysis was used to explore interactions among the DE-FRGs. Hub genes were identified using the random forest algorithm, and their RNA expression levels were validated via RT-qPCR in sham-operated, MCAO model, and acupuncture groups.

RESULTS

The results showed that one hundred twenty-seven DE-FRGs were identified by comparing cerebral ischemic injury tissue with normal brain tissue in mice. KEGG enrichment analysis revealed that these DE-FRGs were primarily involved in the ferroptosis pathway, autophagy-animal pathway, apoptosis pathway, HIF-1 signaling pathway, and longevity-regulating pathway. The top 10 hub DE-FRGs selected through the PPI analysis and random forest algorithm included SLC3A2, FTH1, MAP1LC3A, SLC40A1, TFRC, TMSB4X NRAS CD82 CD44, and PTPN18. Nine genes were confirmed to be significantly differentially expressed between the sham and MCAO mouse models. Moreover, FTH1, SLC40A1, NRAS, CD82, and PTPN18 levels significantly increased after acupuncture in ischemic stroke mice.

CONCLUSION

The ferroptosis pathway, autophagy-animal pathway, apoptosis pathway, HIF-1 signaling pathway, and longevity-regulating pathway were identified as crucial pathways associated with ferroptosis in cerebral ischemic stroke. Bioinformatics analysis and RT-qPCR suggested that FTH1, SLC40A1, NRAS, CD82, and PTPN18 might serve as potential key targets underlying the antiferroptotic effects of acupuncture on ischemic stroke.

摘要

背景

据报道，铁死亡与缺血性脑卒中的发生发展有关；然而，缺乏对铁死亡相关基因的全面研究。在此，通过生物信息学分析和实验，鉴定并验证了缺血性脑卒中中与铁死亡相关的关键差异表达基因（DEGs），并探讨了针刺治疗缺血性脑卒中的潜在机制。

方法

采用高通量RNA测序技术鉴定小鼠脑缺血损伤组织与正常脑组织之间的DEGs。随后，通过将DEGs与铁死亡数据库进行比对，鉴定出差异表达的铁死亡相关基因（DE-FRGs）。进行功能富集分析以研究缺血性脑卒中涉及的潜在信号通路，同时利用蛋白质-蛋白质相互作用（PPI）分析来探索DE-FRGs之间的相互作用。使用随机森林算法鉴定枢纽基因，并通过RT-qPCR在假手术组、大脑中动脉闭塞（MCAO）模型组和针刺组中验证其RNA表达水平。

结果

结果显示，通过比较小鼠脑缺血损伤组织与正常脑组织，鉴定出127个DE-FRGs。KEGG富集分析表明，这些DE-FRGs主要参与铁死亡途径、自噬-动物途径、凋亡途径、缺氧诱导因子-1（HIF-1）信号通路和寿命调节途径。通过PPI分析和随机森林算法筛选出的前10个枢纽DE-FRGs包括溶质载体家族3成员2（SLC3A2）、铁蛋白重链1（FTH1）、微管相关蛋白1轻链3α（MAP1LC3A）、溶质载体家族40成员1（SLC40A1）、转铁蛋白受体1（TFRC）、胸腺素β4（TMSB4X）、神经母细胞瘤RAS病毒癌基因同源物（NRAS）、CD82分子（CD82）、CD44分子（CD44）和蛋白酪氨酸磷酸酶非受体型18（PTPN18）。证实假手术组和MCAO小鼠模型之间有9个基因存在显著差异表达。此外，缺血性脑卒中小鼠针刺后FTH1、SLC40A1、NRAS、CD82和PTPN18的水平显著升高。