Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, China.
Qinghai Key Laboratory of Tibetan Medicine Pharmacology and Safety Evaluation, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining, China.
Zebrafish. 2019 Oct;16(5):434-442. doi: 10.1089/zeb.2018.1722. Epub 2019 Jul 16.
Strategies to reduce reperfusion injury after ischemia have been considered in clinical practice, but few interventions have successfully passed the proof-of-concept stage. In this study, we developed a novel zebrafish larvae hypoxia/reoxygenation (H/R) model to simulate myocardial ischemia/reperfusion injury (MIRI), with potential utility as a drug screening tool. After H/R treatment, videos of transgenic [Tg(cmlc:EGFP)] larval zebrafish hearts were captured using a digital high-speed camera, and the heart rate, diastolic area, systolic area, and total fraction of area changed were quantified. The mRNA expression of and was quantified, and red blood cells (RBCs) were detected by O-dianisidine staining. We found that a decline in cardiac contractility occurred in zebrafish larvae 48 h after hypoxia treatment. Reoxygenation for 2-5 h after 48 h of hypoxia caused heart dysfunction in zebrafish larvae, and were determined to be the optimum conditions for simulating MIRI similar to mammalian models. Our results indicated that heart dysfunction after reoxygenation in zebrafish larvae was accompanied by an upregulated gene expression of a number of myocardial injury biomarkers and increased numbers of RBCs. In conclusion, the novel larval zebrafish H/R model developed in this study could be used for rapid screening and efficacy assessment of MIRI therapeutics.
在临床实践中，人们已经考虑了减少缺血后再灌注损伤的策略，但很少有干预措施成功通过概念验证阶段。在这项研究中，我们开发了一种新颖的斑马鱼幼虫缺氧/复氧 (H/R) 模型，用于模拟心肌缺血/再灌注损伤 (MIRI)，有望成为一种药物筛选工具。H/R 处理后，使用数字高速摄像机拍摄转 [Tg(cmlc:EGFP)] 斑马鱼幼虫心脏的视频，并定量测量心率、舒张面积、收缩面积和总面积变化的分数。定量检测 和 的 mRNA 表达，并通过 O-二苯胺染色检测红细胞 (RBC)。我们发现，缺氧处理 48 小时后，斑马鱼幼虫的心脏收缩力下降。缺氧 48 小时后再复氧 2-5 小时会导致斑马鱼幼虫心脏功能障碍，被确定为模拟类似于哺乳动物模型的 MIRI 的最佳条件。我们的结果表明，再复氧后斑马鱼幼虫心脏功能障碍伴随着许多心肌损伤生物标志物的基因表达上调和 RBC 数量增加。总之，本研究中开发的新型幼虫斑马鱼 H/R 模型可用于快速筛选和评估 MIRI 治疗药物的疗效。
