| 引用本文: |
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[点击复制] |
|
| |
|
|
| 本文已被：浏览 930次 下载 707次 |
| 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 |
| 袁嘉,陈俐彤,刘晓月,李千隽,魏瑢琪,田浩梅,陈楚淘 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 基于不同时间点观察针刺对脑缺血再灌注损伤（CIRI）大鼠缺血半暗带铁死亡相关因子表达的影响，探讨针刺改善CIRI大鼠神经细胞损伤的作用机制。方法 将168只雄性SD大鼠采用随机数字法分为假手术组（42只）和造模组（126只），造模组采用改良Longa线栓法制备大脑中动脉闭塞再灌注（MCAO/R）模型，造模成功后再随机分为模型组、去铁酮（DFP）组和针刺组，每组42只。以上各组再根据取材时间点随机分为24 h组、48 h组和72 h组，每组14只。针刺组捆绑并针刺大椎、百会、水沟穴，每次留针30 min，DFP组灌胃注射DFP溶液（0.125 g/kg），其余各组给予等量0.9%氯化钠溶液灌胃注射，以上各组每隔12 h干预1次，其中24 h组共干预3次，48 h组共干预5次，72 h组共干预7次。干预前后采用改良神经功能缺损评分（mNSS）法评估大鼠神经功能状态，2,3,5-氯化三苯基四氮唑（TTC）染色观察并计算脑梗死体积比，透射电镜观察脑组织缺血半暗带线粒体超微结构，普鲁士蓝染色观察脑组织铁沉积情况，比色法检测脑组织缺血半暗带铁离子（Fe2+）含量，酶联免疫吸附测定（ELISA）检测脑组织缺血半暗带活性氧（ROS）和谷胱甘肽过氧化物酶4（GPX4）含量，硫代巴比妥酸（TBA）及微板法检测脑组织缺血半暗带丙二醛（MDA）、谷胱甘肽（GSH）含量。结果 与假手术组相比，各时间点模型组大鼠mNSS神经功能评分、脑梗死体积比显著升高（P<0.01），各时间点模型组线粒体均发生不同程度的水肿变性，且在72 h呈现典型铁死亡特征、铁沉积数量明显增多，缺血半暗带Fe2+、ROS、MDA含量显著升高（P<0.01），GPX4、GSH含量显著降低（P<0.01）。与模型组比较，各时间点干预组大鼠mNSS神经功能评分、脑梗死体积比显著降低（P<0.01），线粒体铁死亡改变随时间减轻，铁沉积数量明显减少，缺血半暗带Fe2+、ROS、MDA含量降低（P<0.05，P<0.01），GPX4、GSH含量升高（P<0.05，P<0.01）。与DFP组比较，针刺组干预24 h、48 h后ROS含量降低（P<0.05，P<0.01），干预72 h后GPX4含量和干预24 h后MDA含量升高（P<0.05）。结论 针刺对CIRI大鼠具有神经保护作用，其机制可能与调节缺血半暗带铁死亡相关因子、抑制氧化应激损伤、改善神经元铁死亡有关，且随时间呈现动态变化。 |
| 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 |
| DOI：10.3969/j.issn.1674-070X.2025.12.010 |
| 投稿时间：2025-08-03 |
| 基金项目:国家自然科学基金项目（82274662）；湖南省自然科学基金项目（2024JJ5305）；湖南省中医药管理局基金项目（A2024009）。 |
|
| 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 |
| YUAN Jia, CHEN Litong, LIU Xiaoyue, LI Qianjun, WEI Rongqi, TIAN Haomei, CHEN Chutao |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| 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. |
| Key words: cerebral ischemia-reperfusion injury ischemic penumbra acupuncture neurological impairment ferroptosis |
|
二维码（扫一下试试看！） |
|
|
|
|
