湖南中医药大学学报2025，Vol.45Issue(12)：2316-2324,9.DOI:10.3969/j.issn.1674-070X.2025.12.010
基于不同时间点观察针刺对脑缺血再灌注损伤大鼠缺血半暗带铁死亡相关因子表达的影响
Effects of acupuncture on the expression of ferroptosis-related factors in the ischemic penumbra of rats with cerebral ischemia-reperfusion injury at different time points
摘要
Abstract
Objective To explore the mechanism of action of acupuncture in ameliorating neural cell damage in rats with cerebral ischemia-reperfusion injury(CIRI)by observing the effect of acupuncture on the expression of ferroptosis-related factors in the ischemic penumbra of rats with CIRI at different time points.Methods A total of 168 male SD rats were randomly divided into a sham-operated group(n=42)and a modeling group(n=126)using random number tables.The modified Longa suture-occlusion method was used to establish middle cerebral artery occlusion/reperfusion(MCAO/R)models in the modeling group.After successful modeling,the modeling group was further randomly divided into a model group,a deferiprone(DFP)group,and an acupuncture group,with 42 rats in each group.Subsequently,each of these groups was randomly subdivided into 24 h,48 h,and 72 h subgroups based on tissue collection time points,with 14 rats in each subgroup.Rats in the acupuncture group were immobilized and received acupuncture at the"Dazhui(GV14),""Baihui(GV20),"and"Shuigou(GV26)"acupoints,with acupuncture needles left in place for 30 minutes each time.Rats of the DFP group were administered intragastric administration of DFP solution(0.125 g/kg),while rats of the remaining groups received an equivalent volume of 0.9%sodium chloride solution via intragastric administration.All groups were intervened every 12 hours,with the 24 h subgroup receiving three interventions,the 48 h subgroup receiving five interventions,and the 72 h subgroup receiving seven interventions.Neurological function status was assessed using the modified neurological severity score(mNSS)before and after intervention.2,3,5-Triphenyltetrazolium chloride(TTC)staining was performed to observe and calculate the brain infarction volume ratio.Transmission electron microscopy was used to examine the mitochondrial ultrastructure in the ischemic penumbra of the brain tissue.Prussian blue staining was employed to observe the level of iron deposition in the brain tissue.The Fe2+content in the ischemic penumbra of the brain tissue was measured by colorimetry.Enzyme-linked immunosorbent assay(ELISA)was used to determine the content of reactive oxygen species(ROS)and glutathione peroxidase 4(GPX4)in the ischemic penumbra of the brain tissue.The thiobarbituric acid(TBA)method and microplate assay were utilized to measure malondialdehyde(MDA)and glutathione(GSH)content in the ischemic penumbra of the brain tissue.Results Compared with the sham-operated group,rats in the model group at all time points exhibited significantly elevated mNSS neurological function scores(P<0.01),along with a markedly increased brain infarction volume ratio(P<0.01).Mitochondria in the model group showed varying degrees of edema and degeneration at all time points,with typical ferroptosis features and significantly increased iron deposition observed at 72 hours.The contents of Fe2+,ROS,and MDA in the ischemic penumbra significantly increased(P<0.01),while GPX4 and GSH contents significantly decreased(P<0.01).Compared with the model group,rats in the intervention groups at all time points showed significantly lower mNSS neurological function scores(P<0.01),along with a markedly reduced brain infarction volume ratio(P<0.01).Ferroptosis-associated mitochondrial alterations were alleviated over time.The level of iron deposition significantly reduced.The content of Fe2+,ROS,and MDA in the ischemic penumbra decreased(P<0.05,P<0.01),while GPX4 and GSH content increased(P<0.05,P<0.01).Compared with the DFP group,the acupuncture group showed lower ROS content after 24 and 48 hours of intervention(P<0.05,P<0.01),higher GPX4 content after 72 hours,and higher MDA content after 24 hours(P<0.05).Conclusion Acupuncture exerts neuroprotective effects in rats with CIRI.The mechanism may be related to regulating ferroptosis-related factors in the ischemic penumbra,inhibiting oxidative stress damage,and improving neuronal ferroptosis,with dynamic changes over time.关键词
脑缺血再灌注损伤/缺血半暗带/针刺/神经功能缺损/铁死亡Key words
cerebral ischemia-reperfusion injury/ischemic penumbra/acupuncture/neurological impairment/ferroptosis分类
医药卫生引用本文复制引用
袁嘉,陈俐彤,刘晓月,李千隽,魏瑢琪,田浩梅,陈楚淘..基于不同时间点观察针刺对脑缺血再灌注损伤大鼠缺血半暗带铁死亡相关因子表达的影响[J].湖南中医药大学学报,2025,45(12):2316-2324,9.基金项目
国家自然科学基金项目(82274662) （82274662）
湖南省自然科学基金项目(2024JJ5305) （2024JJ5305）
湖南省中医药管理局基金项目(A2024009). （A2024009）
