羟自由基对培养心肌细胞损伤作用的研究

The Study of Damaging Effects of Hydroxyl Radicals on Cultured Myocardial Cells

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 。

Objective: In order to identify the relationship between the accumulation of concentration of free hydroxyl radicals and the injury level of myocardial cell,different concentrations of free hydroxyl radicals were used to act on cultured myocardial cell.Method:Free hydroxyl radicals which were generated by Fenton system were used to act on myocardial cell of suckling mouse for 12h and assay the lactic dehydrogenase(LDH)and malondialhy dride(MDA)in the culture fluid.Assay the activity of myocardial cell by MTT and observe the morphologic changes of myocardial cell by acridine orange staining and transmission electron microscope.Results:Low concentration of free hydroxy radicals in the culture could increase the content of lipid peroxide and decrease enzyme leakage;the mortality of the myocardial cell is low and of mostly apoptosis.With the increasement of the hydroxidion,released LDH and accumulation of the lipid peroxide increase dramatically and cellular mortality raises.But most dead myocardial cells are of were necrosis,with apoptosis cell decreasing.Conclusion:Free hydroxyl radicals,as one kind of active oxygen free radicals,will injure various kinds of myocardial cell.It can induce apoptosis of myocardial cell in low concentration and cause necrosis of myocardial cell.