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Year : 2018  |  Volume : 131  |  Issue : 19  |  Page : 2277-2286

Feasibility Analysis of Oxygen-Glucose Deprivation-Nutrition Resumption on H9c2 Cells In vitro Models of Myocardial Ischemia-Reperfusion Injury

Department of Anesthesiology, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100144, China

Correspondence Address:
Prof. Fu-Shan Xue
Department of Anesthesiology, Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 33 Ba-Da-Chu Road, Shi-Jing-Shan District, Beijing 100144
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/0366-6999.241809

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Background: Oxygen-glucose deprivation-nutrition resumption (OGD-NR) models on H9c2 cells are commonly used in vitro models of simulated myocardial ischemia-reperfusion injury (MIRI), but no study has assessed whether these methods for establishing in vitro models can effectively imitate the characteristics of MIRI in vivo. This experiment was designed to analyze the feasibility of six OGD-NR models of MIRI. Methods: By searching the PubMed database using the keywords “myocardial reperfusion injury H9c2 cells,” we obtained six commonly used OGD-NR in vitro models of MIRI performed on H9c2 cells from more than 400 published papers before January 30, 2017. For each model, control (C), simulated ischemia (SI), and simulated ischemia-reperfusion (SIR) groups were assigned, and cell morphology, lactate dehydrogenase (LDH) release, adenosine triphosphate (ATP) levels, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and inflammatory cytokines were examined to evaluate the characteristics of cell injury. Subsequently, a coculture system of cardiomyocyte-endothelial-macrophage was constructed. The coculture system was dealt with SI and SIR treatments to test the effect on cardiomyocytes survival. Results: For models 1, 2, 3, 4, 5, and 6, SI treatment caused morphological damage to cells, and subsequent SIR treatment did not cause further morphological damage. In the models 1, 2, 3, 4, 5 and 6, LDH release was significantly higher in the SI groups than that in the C group (P < 0.05), and was significantly lower in the SIR groups than that in the SI groups (P < 0.05), except for no significant differences in the LDH release between C, SI and SIR groups in model 6 receiving a 3-h SI treatment. In models 1, 2, 3, 4, 5, and 6, compared with the C group, ATP levels of the SI groups significantly decreased (P < 0.05), ROS levels increased (P < 0.05), and MMP levels decreased (P < 0.05). Compared with the SI group, ATP level of the SIR groups was significantly increased (P < 0.05), and there was no significant ROS production, MMP collapse, and over inflammatory response in the SIR groups. In a coculture system of H9c2 cells-endothelial cells-macrophages, the proportion of viable H9c2 cells in the SIR groups was not reduced compared with the SI groups. Conclusion: All the six OGD-NR models on H9c2 cells in this experiment can not imitate the characteristics of MIRI in vivo and are not suitable for MIRI-related study.


 Abstract in Chinese



背景:H9c2细胞糖氧剥夺-营养恢复(OGD-NR)模型是最常用的模拟在体心肌缺血-再灌注损伤(myocardial ischemia-reperfusion injury (MIRI) 的离体建模方法。但是,至今尚无研究对该模型的可行性进行评估。该实验旨在评价常用的6种OGD-NR模型模拟在体MIRI的可行性。


结果:在模型1、2、3、4、5和6,SI处理可导致细胞形态学损伤,随后实施SIR处理未造成进一步的细胞形态学损伤。除外接受3h SI处理模型6的LDH释放在C、SI和SIR组之间无明显差异之外,模型1, 2, 3, 4, 5和6的SI组LDH释放明显高于C组(P < 0.05),SIR组LDH释放明显低于SI组(P < 0.05)。在模型1、2、3、4、5和6,与C组相比,SI组的ATP明显降低(P<0.05)、ROS生成明显增加(P<0.05)、MMP水平明显降低(P<0.05)。与SI组相比,SIR组的LDH释放明显降低(P<0.05)、ATP水平明显升高(P<0.05)、无大量ROS生成、无MMP坍塌以及无过度的炎症反应。在H9c2细胞-内皮细胞-巨噬细胞共培养系统实验中,与SI组相比,SIR组的活细胞比例无明显降低。


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