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目的 :应用不同浓度的羟自由基作用于培养心肌细胞 ,观察羟自由基浓度的积累与心肌细胞损伤程度的关系。方法 :采用Fenton体系产生羟自由基作用于培养的乳鼠心肌细胞12h ,测定培养液中的乳酸脱氢酶与丙二醛 ,采用MTT法测定心肌细胞活性 ,吖啶橙染色、透射电镜等观察心肌细胞的形态学变化。结果 :低浓度的羟自由基可使培养液中的脂质过氧化物含量增加 ,细胞内酶泄漏较少 ,心肌细胞的死亡率低 ,主要以凋亡为主 ;随着羟离子浓度提高 ,乳酸脱氢酶释放和过氧化脂质积累明显升高 ,细胞死亡率也明显增加 ,但凋亡细胞减少 ,心肌细胞主要以坏死为主。结论 :羟自由基作为一种活性氧自由基 ,依其浓度不同可造成不同类型的心肌细胞损伤 ,低浓度主要诱导心肌细胞凋亡 ,高浓度主要引起心肌细胞坏死
OBJECTIVE: To study the relationship between the concentration of hydroxyl radical and the degree of cardiomyocyte injury in cultured cardiomyocytes with different concentrations of hydroxyl radical. Methods: Hydroxyl radical was induced by Fenton system in cultured neonatal rat cardiomyocytes for 12 hours. Lactate dehydrogenase (MDA) and malondialdehyde (MDA) in culture medium were measured. The activity of cardiomyocytes was measured by MTT assay. Acridine orange staining and transmission electron microscopy Morphological changes of cardiomyocytes. Results: Low concentration of hydroxyl free radicals could increase the content of lipid peroxidation in the medium, less leakage of enzyme in the cells and low mortality of cardiomyocytes, mainly apoptosis; with the increase of the concentration of hydroxyl ions, Lactate dehydrogenase release and lipid peroxidation accumulation was significantly increased, cell death rate was significantly increased, but decreased apoptotic cells, mainly myocardial necrosis. Conclusion: As a kind of reactive oxygen species, hydroxyl radical can cause different types of cardiomyocyte injury according to its concentration. Low concentration mainly induces cardiomyocyte apoptosis, and high concentration mainly causes cardiomyocyte necrosis