上海针灸杂志2025，Vol.44Issue(6)：740-749,10.DOI:10.13460/j.issn.1005-0957.2025.06.0740
基于转录组测序探讨线粒体对缺血性卒中的作用及电针干预机制
Study of the role of mitochondria in ischemic stroke and the mechanism of electroacupuncture intervention based on transcriptome sequencing
摘要
Abstract
Objective To observe the role of mitochondria in ischemic stroke and the action mechanism of electroacupuncture(EA)in prevention and treatment from the perspectives of key targets and signaling pathways.Method Thirty Sprague-Dawley(SD)rats were randomly divided into a control group,a model group,and an intervention group,each consisting of 10 rats.Rat models of ischemic stroke induced by middle cerebral artery occlusion/reperfusion(MCAO/R)were developed in the model and intervention groups using the suture method.The intervention group received EA intervention at Fengchi(GB20)starting from the day of MCAO/R modeling after the operation,and the Bederson score was adopted to assess the neurological deficit after each EA session.When the intervention course finished,cerebral infarction was observed using 2,3,5-triphenyl tetrazolium chloride(TTC)staining;the high-throughput sequencing technology was employed to analyze the differentially expressed genes in rat motor cortex tissues across the three groups.The genes related to mitochondrial structure and function were collected using databases,and the potential target genes were identified by intersecting the differentially expressed genes with the rat mitochondrial structure and function gene dataset.The potential target genes were subjected to KEGG and GO for enrichment analysis using the DAVID online database.Three differentially expressed key target genes were selected using the STRING analysis platform for validation using real-time fluorescence quantitative PCR and Western Blot.Result Compared to the control group,the brain infarction area was significantly larger in the model group rats(P＜0.05)along with a notably increased Bederson score(P＜0.05);compared to the model group,the brain infarction area was significantly smaller in the intervention group(P＜0.05)together with a markedly decreased Bederson score(P＜0.05).In the motor cortex of model group rats,627 differentially expressed genes were identified,and 2 305 differentially expressed genes were identified in the intervention group;22 potential therapeutic targets were obtained by intersecting the differentially-expressed genes with the mitochondrial function gene dataset.The PPI analysis showed that Caspase-1,nucleotide-binding oligomerization domain-like receptor family,pyrin domain-containing 3(Nlrp-3),and Pycard might be the key targets for EA intervention.The KEGG analysis revealed significant enrichment in pathways related to NOD-like receptor signaling,cell-soluble DNA sensing,and necrosis.The GO enrichment results indicated that the biological process in acupuncture treatment of ischemic stroke through mitochondria-related genes were mainly associated with apoptosis,inflammatory responses,and immune responses.Cellular components mainly involved mitochondria,cytoplasm,endoplasmic reticulum,and inflammasomes.Molecular functions involved protein binding,carbohydrate binding,and signal adapter activities.Compared to the control group,the model group showed elevated relative expression levels of Caspase-1,Nlrp-3,and Pycard mRNAs and proteins(P＜0.05);compared to the model group,the intervention group exhibited reduced relative expression levels of Caspase-1,Nlrp-3,and Pycard proteins(P＜0.05)and significantly decreased Nlrp-3 and Pycard mRNA expression levels(P＜0.05).Conclusion The mechanism of EA in treating ischemic stroke exhibits characteristics of multiple components,multiple targets,and multiple pathways;EA may modulate mitochondrial function to promote the recovery of motor neurons and reduce the cerebral infarction area in ischemic stroke rats by down-regulating the expression of Caspase-1,Nlrp-3,and Pycard mRNAs and proteins in the ischemic penumbra of rat brain tissue.关键词
针刺疗法/电针/脑缺血/卒中/缺血再灌注/转录组测序/富集分析/大鼠Key words
Acupuncture therapy/Electroacupuncture/Cerebral ischemia/Stroke/Ischemia-reperfusion/Transcriptome sequencing/Enrichment analysis/Rats分类
医药卫生引用本文复制引用
郝铃钰,张英杰,陈吕佳,徐鸣曙..基于转录组测序探讨线粒体对缺血性卒中的作用及电针干预机制[J].上海针灸杂志,2025,44(6):740-749,10.基金项目
上海市卫生健康委员会科研项目(2020LQ020) （2020LQ020）
上海市科学技术委员会科研计划项目(17ZR1427500) （17ZR1427500）
上海市人才发展基金项目(2017094) （2017094）
