论文部分内容阅读
目的:建立心肌细胞缺血再灌注模型,有助于在细胞层面对缺血再灌注损伤的保护机制进行探讨,防治临床的无复流现象。方法:分离并纯化乳鼠心肌细胞,培养心肌细胞至完全同步化。以高纯氮气和纯氧分别模拟缺血液和再灌注液。预设系列缺血时间和再灌注时间,测定细胞活力指标制作时间一细胞存活率曲线图,寻求最佳缺血再灌注时间窗。结果:心肌细胞培养第5天搏动呈同步化,频率约90次/min。与对照组相比,缺血5 h损伤组与其他组的差异有统计学意义(P<0.05)。与对照组相比,再灌注2、4和6 h心肌细胞存活率的下降都有统计学差异,P值分别为0.013、0.009和0.009。再灌注时间-细胞存活率曲线图提示再灌注2 h曲线斜率最大,存活率下降最显著。结论:确立心肌细胞缺血再灌注模型缺血时间5 h。再灌注时间2 h。
OBJECTIVE: To establish a cardiomyocyte model of ischemia-reperfusion, which is helpful to explore the protective mechanism of ischemia-reperfusion injury at the cellular level and to prevent and cure the clinical no-reflow phenomenon. Methods: Isolated and purified neonatal rat cardiomyocytes were cultured for complete synchronization. High-purity nitrogen and pure oxygen were used to simulate ischemia and reperfusion fluid respectively. Pre-set series of ischemia time and reperfusion time, determination of cell viability indicators of a cell survival curve, seek the best time window of ischemia-reperfusion. Results: Cardiomyocytes were pulsatile on day 5 with a frequency of about 90 beats / min. Compared with the control group, the difference between the ischemic 5 h group and other groups was statistically significant (P <0.05). Compared with the control group, there was a significant difference in the survival rate of cardiomyocytes at 2, 4 and 6 hours after reperfusion, with P values of 0.013, 0.009 and 0.009, respectively. The curve of reperfusion time and cell survival rate showed that the curve of 2 h after reperfusion had the largest slope and the most significant decrease in the survival rate. Conclusion: The ischemia time of cardiomyocyte ischemia-reperfusion model was established for 5 h. Reperfusion time 2 h.