刺血醒脑法对大鼠脑缺血再灌注损伤的脑保护作用实验研究  

Exploring the Neuroprotective Mechanism of Blood-Letting Therapy on Cerebral Ischemia-reperfusion Injury in Rats Based on Transcriptomics

作　　者：曾子修 刘晓君 官玥玥 冉丽 罗晓琼 唐军[1] Zeng Zixiu;Liu Xiaojun;Guan Yueyue;Ran Li;Luo Xiaoqiong;Tang Jun(Chongqing Traditional Chinese Medicine Hospital,Chongqing 400021,China)

机构地区：[1]重庆市中医院,重庆400021

出　　处：《中国中医急症》2025年第3期409-415,共7页Journal of Emergency in Traditional Chinese Medicine

基　　金：2022年度重庆英才计划“包干制”项目(cstc2022ycjh-bgzxm0138);重庆市巴渝岐黄学者支持项目(渝中医[2023]23号)。

摘　　要：目的利用转录组测序(RNA-seq)技术探究刺血醒脑法对大鼠脑缺血再灌注损伤的脑保护作用及其潜在机制。方法将75只SD大鼠随机分为假手术组、模型组和刺血醒脑组。采用改良线栓法建立脑缺血再灌注(MCAO/R)模型,并对模型组大鼠实施刺血醒脑法干预。在缺血再灌注后0、24、48、72 h关键时间点,记录大鼠存活率和体重变化,采用Garcia评分评估神经功能,旷场试验评价行为学表现,TTC染色法观察和定量分析脑梗死体积,HE染色检测脑组织病理形态学,TUNEL染色检测细胞凋亡;利用RNA-seq技术筛选刺血醒脑法治疗后的关键靶点及调控途径,并通过Western blotting验证相关通路。结果在脑缺血再灌注后72 h,刺血醒脑法显著改善了MCAO/R模型大鼠的神经功能缺损及行为学测试结果,缩小了脑梗死体积,保护了脑组织细胞结构的完整性,并减少了神经细胞凋亡(P<0.05);转录组学研究显示,刺血醒脑法干预后,差异基因富集通路与NOD样受体信号通路显著相关,其中Nlrp3、Casp1、IL-18等基因表达显著下调。Western blotting结果表明,与模型组相比,刺血醒脑组中NLRP3、ASC、Caspase-1、IL-1β、IL-18等蛋白表达水平明显下降(P<0.05)。结论刺血醒脑法对脑缺血再灌注损伤的大鼠具有显著的脑保护作用,其作用机制可能涉及抑制NLRP3/Caspase-1/IL-1β信号通路,进而发挥抗炎效应。Objective:To investigate the neuroprotective effects of blood-letting therapy on cerebral ischemiareperfusion injury in rats and to explore its underlying mechanisms using RNA sequencing(RNA-seq)technology.Methods:A total of 75 SD rats were randomly divided into three groups:a sham surgery group,a model group,and a blood-letting therapy group.A modified thread-occlusion method was used to establish a Middle Cerebral Artery Occlusion and Reperfusion(MCAO/R)model,and blood-letting therapy was applied to the model group.At key time points(0,24,48,and 72 hours)after ischemia-reperfusion,survival rates and body weight changes were recorded.Neurological function was assessed using the Garcia score,behavioral performance was evaluated using the open field test,and brain infarct volume was observed and quantified using TTC staining.Histopathological morphology of brain tissue was examined with HE staining,and cell apoptosis was detected with TUNEL staining.RNA-seq technology was utilized to identify key targets and regulatory pathways after treatment with blood-letting therapy,and Western blotting was employed to validate the related pathways.Results:At 72 hours after ischemia reperfusion,the blood-letting therapy significantly improved neurological deficits and behavioral test results in MCAO/R model rats,reduced brain infarct volume,preserved the integrity of brain tissue cellular structure,and decreased neuronal apoptosis(P<0.05).Transcriptomic analysis revealed that the differentially expressed genes enriched in pathways significantly associated with the NOD-like receptor signaling pathway were downregulated,including Nlrp3,Casp1,and IL-18.Western blotting results indicated that the expression levels of NLRP3,ASC,Caspase-1,IL-1β,and IL-18 proteins were significantly decreased in the blood-letting therapy group compared to the model group(P<0.05).Conclusion:Blood-letting therapy exerts significant neuroprotection in rats with cerebral ischemia-reperfusion injury,and its mechanism of action may involve the inhibition

关 键 词：脑缺血再灌注 针刺 刺血醒脑法 转录组学 NLRP3/Caspase-1/IL-1β 大鼠 

分 类 号：R743.3[医药卫生—神经病学与精神病学]