| 游晓华,董斐斐,李松华,刘夙璇,赵仙先.孤儿核受体Nur77对缺/复氧损伤中心肌细胞自噬的调节[J].现代生物医学进展英文版,2018,(2):219-221. |
| 孤儿核受体Nur77对缺/复氧损伤中心肌细胞自噬的调节 |
| Regulatory Role of Orphan Nuclear Receptor Nur77 in Cardiomyocytes Autophagy during Hypoxia/Reoxygenation Injury |
| Received:June 16, 2017 Revised:July 12, 2017 |
| DOI：10.13241/j.cnki.pmb.2018.02.005 |
| 中文关键词: 孤儿核受体77 心肌细胞 缺复氧损伤 自噬 |
| 英文关键词: Nur77 Cardiomyocytes Hypoxia/reoxygenation injury Autophagy |
| 基金项目:上海市自然科学基金项目（14ZR1408100） |
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| 中文摘要: |
| 摘要 目的：研究孤儿核受体Nur77对缺/复氧损伤中心肌细胞自噬的调节作用。方法：差速贴壁法分离乳鼠心肌细胞，经免疫荧光染色鉴定纯度。缺氧（1 %O2、5 %CO2和94 %N2）培养12 h后，常氧培养2 h构建心肌细胞缺/复氧损伤。实时定量PCR和western blot的方法检测Nur77的表达变化。通过siRNA转染抑制心肌细胞nur77表达，通过自噬标志蛋白表达改变作为细胞自噬水平的变化。结果：原代分离的心肌细胞纯度95 %以上。缺氧12 h和缺/复氧（12 h/2 h）刺激后，心肌细胞中Nur77表达都明显升高（P<0.01）。与缺氧组相比，缺/复氧组细胞质中的水平明显增加（P<0.01），细胞核中Nur77水平无明显变化。抑制Nur77后，缺/复氧组自噬水平明显降低，缺氧组心肌细胞自噬水平无明显变化。结论：Nur77参与缺/复氧损伤中心肌细胞自噬水平的调节。 |
| 英文摘要: |
| ABSTRACT Objective: To study regulation of orphan nuclear receptor Nur77 on cardiomyocytes autophagy during hypoxia/reoxygenation injury. Methods: Primary neonatal rat cardiomyocytes were obtained by differential adherence method, and were identified purity by immunofluorescence staining. Cardiomyocytes hypoxia/reoxygenation injury was established by culturing in hypoxic condition (1% O2, 5% CO2 and 94% N2) for 12 h, followed by reoxygenation for 2 h. Expression of nur77 was determined by real-time PCR. Cardiomyocytes Nur77 expression was inhibited by siRNA transfection. Autophagy level was determined according to the expression of autophagy marker. Results: The purity of separated cardiomyocytes reached by more than 95 %. The expression of Nur77 was significantly increased in cardiomyocytes after hypoxia (12 h) and hypoxia/reoxygenation (12 h/2 h) treatment (P<0.01). Compared with hypoxia group, Nur77 was detected significantly increase in cytoplasm from hypoxia/reoxygenation treated cardiomyocytes, but no changes in nucleus. By Nur77 inhibition, the autophagy level was significantly decreased in hypoxia/reoxygenation treated cardiomyocytes, but no significantly change was found in hypoxia treated cardiomyocytes. Conclusion: Nur77 was involved in regulation of cardiomyocytes autophagy during hypoxia/reoxygenation injury. |
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