| 王 艳,宗 媛,赵 品,姜 静,姚立农,陶 蕾.大鼠心肌缺血再灌注损伤模型线粒体自噬变化的研究[J].现代生物医学进展英文版,2018,(13):2452-2456. |
| 大鼠心肌缺血再灌注损伤模型线粒体自噬变化的研究 |
| Mitophagy Changes in Rat Model during Myocardial Ischemia-reperfusion Injury |
| Received:October 24, 2017 Revised:November 19, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.13.010 |
| 中文关键词: 缺血-再灌注 线粒体 线粒体自噬 微管相关蛋白轻链3（LC3） PINK1 Parkin |
| 英文关键词: Ischemia-reperfusion Mitophagy Mitochondria LC3 PINK1 Parkin |
| 基金项目:国家自然科学基金项目（81470414） |
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| 中文摘要: |
| 摘要 目的：观察大鼠心肌缺血再灌注损伤模型不同时间点线粒体及线粒体自噬的变化。方法：成年雄性SD 大鼠40 只，随机分为假手术对照组（sham 组）：开胸不进行冠状动脉左前降支（Left anterior descending coronary artery, LAD）血流阻断；缺血再灌注组2 h组（I/R 2 h组）、24 h组（I/R 24 h组）及48 h组（I/R 48 h组），以上3组均阻断LAD 30 min，分别于再灌注后2 h、24 h、48 h观察心肌ATP含量，线粒体膜电位水平变化，透射电镜下观察线粒体及线粒体自噬超微结构变化，western blot法测定线粒体自噬蛋白PINK1、Parkin、p62、LC3B及线粒体膜蛋白Tom20表达水平。结果：与对照组相比，线粒体膜电位水平及心肌组织ATP 含量于再灌注2 h开始下降，24 h下降最显著，48 h有所改善，线粒体超微结构损伤再灌注24 h最为明显，48 h有所改善。PINK1、Parkin、p62蛋白表达于损伤后2 h增强，于再灌注后24 h升高最显著，持续至48 h，LC3BⅡ表达于损伤后24 h增强，同样持续至48 h。透射电镜下可见线粒体自噬体于再灌注后24 h明显增多，并持续至48 h。结论：大鼠心肌缺血再灌注损伤后，线粒体功能与形态损伤以损伤后24 h最为显著，至损伤后48 h后好转；线粒体自噬水平升高以损伤后24 h最为显著，且维持至损伤后48 h，提示两者之间可能存在关联。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate mitochondria and mitophagy changes at different indicated time during myocardial ischemia reperfusion in rat model. Methods: Forty male SD rats were randomly divided into sham operation group, 2 h after ischemia reperfusion injury group (I/R 2h,), 24 h after ischemia reperfusion injury group (I/R 24h), 48 h after ischemia reperfusion injury group (I/R 48 h). Myocardial ischemia reperfusion injury model was induced by ligating anterior descending branch of coronary artery. Sham operation group was treated identically except left anterior descending was not tied. The other three groups were treated with 30 min of occlusion and reperfusion of 2 h, 24 h, or 48 h, respectively. The level of myocardium ATP and transmembrane potential of mitochondrion were detected. Ultra-structure changes of mitochondria and mitophagy were observed under electron microscope. The expression of mitophagy related proteins such as PINK1, Parkin, p62 and LC3B were measured by western blotting, as well as the expression of mitochondrial membrane protein Tom20. Results: Compared with sham group, the content of myocardium ATP and transmembrane potential of mito- chondrion decreased in I/R 2 h group, the lowest value of which were in I/R 24 h group(P<0.05). However, the ATP and the transmem- brane potential of mitochondrion were elevated in I/R 48 h group(P<0.05). The level of mitophagy related proteins such as PINK1, Parkin, p62 increased in I/R 2h group and last till 48 h after I/R(P<0.05). The expression of LC3BⅡprotein enhanced in 24 h group and last till 48 h after I/R(P<0.05). Obviously ultra-structure changes of mitochondria and mitophagy were observed under electron micro- scope. Conclusion: The damages of mitochondrial was observed at 24 h after myocardial ischemia reperfusion injury, while elevated levels of mitophagy were detected from 24 h to 48 h after myocardial ischemia reperfusion injury, suggesting a relationship between to the changes of mitochondria and autophagy. |
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