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原代培养5天的乳大鼠心肌细胞,用无钙及不含血清的培养基孵育60分钟,在细胞内钙耗竭后加入氯化钙(使钙浓度达2.5mM/ml)造成细胞钙矛盾损伤。实验完毕,按细胞化学染色方法显示各组心肌细胞的线粒体标志酶——琥珀酸脱氢酶。用MPV-3型显微分光光度计测量细胞的琥珀酸脱氢酶染色灰度(它代表了该酶活性的强度)和细胞表面积。结果显示,缺钙孵育时,心肌细胞琥珀酸脱氢酶活性(平均灰度为666.58)及细胞表面积(平均为1743.23)分别与对照组(平均灰度668.12,平均表面积为1789.19)相比(P>0.05),差异无显著性;补钙孵育60分钟后,心肌细胞的琥珀酸脱氢酶活性降低(平均灰度为498.92),细胞极度孪缩,表面积显著减少(平均为1449.14),和对照组和无钙组相比(P<0.01),差异有高度显著性。实验表明钙矛盾损伤中,细胞有极度孪缩和线粒体琥珀酸脱氢酶的损伤变化。
Primary cultured rat cardiac myocytes for 5 days were incubated in calcium-free and serum-free media for 60 min. Ca2 + (Ca2 + up to 2.5 mM / ml) was added after intracellular calcium depletion resulted in calcium-cell contradiction damage. After the experiment, the mitochondrial marker enzyme succinate dehydrogenase of cardiomyocytes in each group was shown by cytochemical staining. The cell’s succinate dehydrogenase staining intensity (which represents the strength of the enzyme activity) and the cell surface area were measured using a MPV-3 spectrophotometer. The results showed that the activity of succinate dehydrogenase (average gray degree 666.58) and cell surface area (average 1743.23) of cardiomyocytes in calcium-deficient group were significantly lower than those in control group (mean gray value 668.12, average surface area 1789.19) > 0.05). There was no significant difference between the two groups (P> 0.05). After 60 minutes of calcium supplementation, the activity of succinate dehydrogenase in cardiomyocytes was decreased (mean gray value was 498.92), cells were extremely twinned and the surface area was significantly reduced (average 1449.14) Compared with no calcium group (P <0.01), the difference was highly significant. Experiments show that calcium contradictory damage, the cells have extreme twins and mitochondrial succinate dehydrogenase damage changes.