摘要
目的探索一种快速建立心肌细胞缺氧/复氧损伤模型的方法。方法取新生1d^3d SD乳鼠,原代心肌细胞培养,观察连二亚硫酸钠(Na_2S_2O_4)不同浓度(1,2,4mmol/L)分别缺氧1h/复氧24h及在4mmol/LNa_2S_2O_4的终浓度下,缺氧30min,观察不同复氧时间(1h,2h,3h,4h,5h,6h),利用MTT法检测心肌细胞存活率及化学比色法测定乳酸脱氢酶(LDH)的释放,并进行细胞形态学观察。结果乳鼠心肌细胞给予不同浓度Na_2S_2O_4进行缺氧1h/复氧24h处理后,发现Na_2S_2O_4浓度为4mmol/L时,MTT的OD值与正常对照组相比有显著性差异,同时上清液中LDH含量升高,差异具有统计学意义(P<0.01)。在Na_2S_2O_4终浓度为4mmol/L的基础上,给予缺氧30min,复氧1h^6h,心肌细胞存活率显著降低,上清液中LDH释放显著增加,差异具有统计学意义(P<0.01)。结论使用Na_2S_2O_4建立乳鼠心肌细胞缺氧/复氧损伤模型方便、快捷、高效。
Objective To explore a rapid method of establishing cardiomyocyte hypoxia/reoxygenation injury model. Methods The primary cultured cardiomyocytes were cultured with newborn SD rats. （1,2, 4 mmol/L） were used to observe the effects of sodium dithionite （Na2S2O4） on hypoxia for lh and reoxygenation for 24 hours. The final concentration of 4mmol/LNa2S2O4 in myocardial cells, after 30 min of hypoxia reoxygenation in different observation time, detection of myocardial cell survival rate was measured by lactate dehydrogenase and chemical colorimetric method using the method of MTT （LDH） release, and the morphology of ceils was observed. Results The OD value of MTF was significantly different from that of normal control group when Na2S2O4 concentration was greater than 4mmol/L, and the content of LDH in supernatant was significantly higher than that in control group （P 〈 0.05） , and the difference was statistically significant （P 〈 0.01 ）. On the basis of Na2S2O4 final concentration of 4mmol/L, hypoxia 30min, reoxygenation 1h～ 6h, cardiomyocyte survival rate decreased significantly, supernatant LDH release increased significantly, the difference was statistically significant （P 〈 0.01 ）. Conclusion The use of Na2S2O4 to establish neonatal rat cardiomyocytes in hypoxia/reoxygenation injury model is convenient, fast and efficient.
出处
《牡丹江医学院学报》
2017年第2期1-5,共5页
Journal of Mudanjiang Medical University
