高血糖条件下NF-κB激活对小鼠心肌缺血再灌注损伤的影响及其机制  

Effect and mechanism of NF-κB activation on myocardial ischemia-reperfusion injury under hyperglycemic condition

作　　者：张宇[1] 阿布都赛米·艾尼 多力昆·木台力甫 郑博源 阿布都乃比·麦麦提艾力[1] ZHANG Yu;Abdusami Ayni;Dolikun Muta'ali;ZHENG Boyuan;Abudunai Meimetiali(Department of Cardiac Surgery,The First Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,China)

机构地区：[1]新疆医科大学第一附属医院心脏外科,乌鲁木齐830011

出　　处：《山东医药》2024年第25期1-5,共5页Shandong Medical Journal

基　　金：国家自然科学基金资助项目(82060907)。

摘　　要：目的探讨高血糖条件下核因子κB(NF-κB)激活对小鼠心肌缺血再灌注(IR)损伤的影响及其机制。方法选择雄性C57BL/6J小鼠48只,适应性饲养1周,随机分为NG-Sham组(非DM+假手术)与NG-IR组(非DM+IR)、DM-Sham组(DM+假手术)与DM-IR组(DM+IR)以及DM-IR-NC组(DM+IR+阴性对照)与DM-IR-NF-κB组(DM+IR+NF-κB),每组8只。NG-Sham组与NG-IR组正常饲料喂养,NG-IR组通过结扎冠状动脉左前降支构建IR模型(缺血60 min再灌注24 h),NG-Sham组开胸暴露冠状动脉左前降支,但不结扎。DM-IR组连续4天腹腔注射链脲佐菌素40 mg/kg诱导DM模型,成模后1周同法构建IR模型。DM-Sham组同法诱导DM模型,成模后1周开胸暴露冠状动脉左前降支,但不结扎。DM-IR-NF-κB组与DM-IR-NC组同法诱导DM模型,成模后1周同法构建IR模型,DM-IR-NF-κB组开胸前30 min尾静脉注射NF-κB重组蛋白0.5 mg/kg,DM-IR-NC组开胸前30 min尾静脉注射等量溶剂。采用TTC染色法评估NG-IR组与DM-IR组心肌缺血面积,采用RT-qPCR法检测NG-IR组与DM-IR组心肌组织NF-κB、IL-6、IL-1β、TNF-αmRNA相对表达量。采用JC-1染色法评估NG-Sham组、NG-IR组、DM-Sham组和DM-IR组心肌细胞线粒体膜电位,采用MitoSOX染色法评估心肌细胞线粒体活性氧(ROS)含量。采用Western blotting法检测NG-IR组与DM-IR组心肌组织AMPK、p-AMPK蛋白相对表达量,计算p-AMPK/AMPK。采用比色法检测DM-IR-NC组与DM-IR-NF-κB组血清AST、LDH含量,采用双抗体夹心ELISA法检测血清CK-MB含量。结果与NG-IR组比较,DM-IR组心肌缺血面积增加(P<0.05),心肌组织NF-κB mRNA相对表达量上调(P均<0.05),而心肌组织IL-6、IL-1β、TNF-αmRNA相对表达量变化不明显(P均>0.05)。与NG-Sham组比较,NG-IR组与DM-Sham组心肌细胞线粒体膜电位下降,心肌细胞线粒体ROS含量升高(P均<0.05);与NG-IR组比较,DM-IR组心肌细胞线粒体膜电位下降,心肌细胞线粒体ROS含量升高(P均<0.05)。Pearson相关分析发现,IR小鼠心肌组织NF-κObjective To investigate the effect and mechanism of the activation of nuclear factorκB(NF-κB)on myocardial ischemia-reperfusion(IR)injury under hyperglycemic condition.Methods Forty-eight male C57BL/6J mice were acclimated for one week and were randomly assigned into six groups:non-diabetic sham-operated group(NG-Sham group),non-diabetic ischemia-reperfusion group(NG-IR group),diabetic sham-operated group(DM-Sham group),diabetic ischemia-reperfusion group(DM-IR group),diabetic ischemia-reperfusion negative control group(DM-IR-NC group),and diabetic ischemia-reperfusion NF-κB recombinant protein intervention group(DM-IR-NF-κB group),with eight mice in each group.Mice in the NG-Sham and NG-IR groups were fed the normal diet.Mice in the NG-IR group underwent myocardial infarction(MI)modeling by ligation of the left anterior descending(LAD)coronary artery,while mice in the NG-Sham group had the LAD artery exposed but not ligated.Mice in the DM-IR group received intraperi‑toneal injections of streptozotocin(STZ)at 40 mg/kg for four consecutive days to induce diabetes mellitus(DM).One week after the establishment of the DM model,MI was induced by LAD ligation.Mice in the DM-Sham group underwent the same procedure for DM induction and had the LAD artery exposed but not ligated.Mice in the DM-IR-NF-κB and DM-IR-NC groups underwent the same DM induction procedure and MI modeling.Mice in the DM-IR-NF-κB group received an intravenous injection of NF-κB recombinant protein at 0.5 mg/kg via the tail vein 30 minutes before chest opening,while mice in the DM-IR-NC group did not receive the NF-κB recombinant protein.The myocardial infarct area in the NG-IR and DM-IR groups was assessed by TTC staining.The mRNA expression levels of NF-κB,IL-6,IL-1β,and TNF-αin the myocardial tissues were measured by RT-qPCR.The mitochondrial membrane potential in the NG-Sham,NG-IR,DM-Sham,and DM-IR groups was evaluated using JC-1 staining,and mitochondrial reactive oxygen species(ROS)content was assessed using MitoSOX staining.AMPK

关 键 词：心肌缺血再灌注损伤 高血糖 核因子ΚB 腺苷酸活化蛋白激酶 线粒体质量 小鼠 

分 类 号：R654[医药卫生—外科学]