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目的丙烯醛是一种具有高反应性的不饱和脂肪醛,它是丙烯胺的代谢产物,在体内外对各种细胞具有毒性作用。本研究探讨丙烯醛对成年小鼠心肌细胞氧自由基及钙浓度的影响及细胞凋亡的作用。方法采用改良langendorff方法分离成年小鼠心肌细胞。分别应用DCF及Fura-2 AM测定细胞内氧自由基水平及钙离子钙浓度,WST法测定心肌细胞存活率,观察凋亡DNA片段及细胞凋亡的形态变化。结果应用1μmol/L丙烯醛可明显增加心肌细胞内氧自由基水平及钙离子浓度,分别达到用药前的12倍及2倍。而且,丙烯醛引起心肌细胞损伤是一种剂量依赖性的,应用25,50及100μmol/L丙烯醛治疗的心肌细胞存活率明显低于对照组(P<0.01)。丙烯醛处理心肌细胞后,可观察到DNA断裂片段及典型的凋亡细胞形态变化。结论丙烯醛可引起成年小鼠心肌细胞的凋亡,其作用可能是由于细胞内氧自由及钙浓度的增加。
Purpose Acrolein is a highly reactive, unsaturated fatty aldehyde that is the metabolite of acrylamides and has toxic effects on various cells both in vitro and in vivo. This study was designed to investigate the effects of acrolein on oxygen free radicals and calcium concentration in adult mouse cardiomyocytes and the role of apoptosis. Methods Adult mouse cardiomyocytes were isolated by modified langendorff method. DCF and Fura-2 AM were used to determine the level of intracellular oxygen free radicals and calcium ion concentration, WST method was used to determine the survival rate of cardiomyocytes, morphological changes of apoptotic DNA fragments and apoptosis were observed. Results Application of 1μmol / L acrolein significantly increased oxygen free radical levels and calcium concentration in cardiomyocytes, reaching 12 and 2 times before administration. Moreover, acrolein-induced cardiomyocyte injury was dose-dependent. The viability of cardiomyocytes treated with 25, 50 and 100 μmol / L acrolein was significantly lower than that of the control group (P <0.01). After acrolein treatment of cardiomyocytes, the morphological changes of DNA fragments and typical apoptotic cells were observed. Conclusion Acrolein can induce cardiomyocyte apoptosis in adult mice, which may be due to the increase of intracellular free oxygen and calcium concentration.