EMRE在小鼠心肌缺血损伤中的作用及其相关机制的研究

发布时间：2019-02-26 08:08

【摘要】：目的:研究调节线粒体钙单向转运体活性所必须的蛋白(essential MCU regulator,EMRE)在小鼠心肌缺血损伤中发挥的作用及其相关机制。方法:利用心肌点注射EMRE腺病毒(Ad-EMRE)上调心肌组织的EMRE分子表达水平,对照小鼠心肌点注射增强绿色荧光蛋白腺病毒(Ad-eGEP),48 h后,采用结扎小鼠左冠状动脉前降支方法建立心肌梗死(MI)模型。实验设置三组:假手术组(给予Ad-eGFP);心肌梗死对照组(给予Ad-eGFP);心肌梗死处理组(给予Ad-EMRE)。三周后,利用小动物超声系统测定小鼠心脏功能;Western blot及免疫组化检测EMRE分子的表达;麦胚凝集素(WGA)染色检测心肌细胞肥大程度;马松(MASSON)染色检测心肌组织胶原含量;原位末端标记法(TUNEL)检测心肌组织的细胞凋亡;Western blot检测心肌组织半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和半胱氨酸天冬氨酸蛋白酶-9(Caspase-9)的表达;MitoSOX荧光探针检测线粒体ROS水平。结果:与心肌梗死对照组相比,心肌梗死处理组的心功能明显变差,心肌肥厚明显加重,心肌组织胶原纤维含量更高。另外,心肌梗死处理组的心肌细胞凋亡明显增多,Caspase-3和Caspase-9表达明显升高,线粒体活性氧簇产生也明显增多。结论:EMRE过表达可增加线粒体ROS生成,诱导心肌细胞凋亡,加重心肌缺血损伤。
[Abstract]:Aim: to study the role and mechanism of mitochondrial calcium uniporter protein (essential MCU regulator,EMRE in myocardial ischemia injury in mice. Methods: EMRE adenovirus (Ad-EMRE) was injected into myocardium to up-regulate the expression of EMRE molecule in myocardium, and green fluorescent protein adenoviruses (Ad-eGEP) were injected into the myocardium of control mice. 48 h later, the expression level of EMRE was up-regulated. (MI) model of myocardial infarction was established by ligating left anterior descending coronary artery in mice. Three groups were divided into three groups: sham operation group (Ad-eGFP), myocardial infarction control group (Ad-eGFP), myocardial infarction treatment group (Ad-EMRE). Three weeks later, the expression of cardiac function; Western blot, EMRE molecule, wheat germ lectin (WGA) staining, collagen content in myocardium and collagen content in myocardium were detected by small animal ultrasound system, wheat germ lectin (WGA) staining and Ma Song (MASSON) staining, respectively. The expression of caspase-3 (Caspase-3) and caspase-9 (Caspase-9) in myocardial tissue was detected by in situ end labeling (TUNEL), and the expression of caspase-3 and Caspase-9 was detected by; Western blot. Mitochondrial ROS level was detected by MitoSOX fluorescence probe. Results: compared with the control group, the cardiac function, myocardial hypertrophy and collagen fiber content in the myocardial infarction group were significantly worse and higher than those in the control group. In addition, myocardial apoptosis, Caspase-3 and Caspase-9 expression and mitochondrial active oxygen cluster production were significantly increased in myocardial infarction group. Conclusion: overexpression of EMRE can increase mitochondrial ROS production, induce cardiomyocyte apoptosis and aggravate myocardial ischemia injury.
【作者单位】：中国人民解放军第四军医大学第一附属医院心血管内科;空军94195部队卫生队;解放军兰州总医院心内科;宝鸡市第二中医医院呼吸内分泌科;中国人民解放军第四军医大学唐都医院骨科;西安市中心医院;
【基金】：陕西省自然科学基础研究计划资助项目(2015JM8479)
【分类号】：R54

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