建立新生大鼠心肌细胞缺氧／复氧模型

目的：探讨缺氧密闭法建立新生SD大鼠心肌细胞缺氧／复氧模型的可行性与稳定性。方法：将体外培养的新生SD大鼠心肌细胞随机分为对照组（不做处理）和实验组（进行不同时间（30min、1-4h）的缺氧／复氧处理）。DCFH-DA荧光探针检测活性氧（ROS）浓度；AO／EB双染法及AnexinV／PI流式细胞术检测细胞凋亡率。结果：对照组ROS浓度及细胞凋亡率均较低；而实验组随着缺氧时间的延长，ROS浓度逐渐增加，细胞凋亡数逐渐增多。不同缺氧时间段细胞凋亡率与对照组比，除缺氧30min组外，1-4h组差异均有统计学意义（P〈0．05，P〈0．01）。结论：厌氧产气袋．缺氧密闭系统建立新生SD大鼠心肌细胞缺氧／复氧模型稳定、可行且重复性好。

Establishment of Hypoxia/Reoxygenation Model for Cardiomyocytes in Neonatal Rat

Objective： To investigate the practicability and stability of establishing hypoxia/reoxygenation model for cardiomyocytes in neonatal rat with an anaero-pack system. Methods： The cardiomyocytes in rats were divided into control group（no processing）and experimental group（different stages of hypoxia/reoxygenation）. The reactive oxygen species （ROS） content was detected by DCFH-DA fluorescence probe. The cells apoptosis was determined by AO/EB double staining and flow cytometry. Results： The ROS content and apoptosis rate in the control group were low. With the prolonging of hypoxia time, the ROS content and apoptosis rate were gradually increasing in the experimental group. Compared with the control group, the apoptosis rate in the experimental group was significantly increased（P〈0.05, P〈0.01）, excluding the 30 minutes group. Conclusion： The establishment of the hypoxia/reoxygenation model for cardiomyocytes in neonatal rat with an anaero-pack system is stable, practicable and repeatable.