摘要
Background： Oxygen-glucose deprivation-nutrition resumption （OGD-NR） models on H9c2 cells are commonly used in vitro models of simulated myocardial ischemia-reperfusion injury （M1RI）, 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 coeulture system was dealt with SI and SIR treatments to test the effect on cardiomyocytes survival. Results： For models l, 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 tbr 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 S1 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 MIRl-related study.
Background： Oxygen-glucose deprivation-nutrition resumption （OGD-NR） models on H9c2 cells are commonly used in vitro models of simulated myocardial ischemia-reperfusion injury （M1RI）, 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 coeulture system was dealt with SI and SIR treatments to test the effect on cardiomyocytes survival. Results： For models l, 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 tbr 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 S1 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 MIRl-related study.
基金
The work was supported by grants from the major projects of Scientific and Research Found of Plastic Surgery Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College （No. Z2017001） and the National Natural Science Foundation of China （No. 81470019）.
