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目的 :探讨bcl 2基因转染对热应激心肌细胞保护作用的机制。方法 :分离、培养乳鼠心肌细胞 ,用脂质体转染法将bcl 2基因转染入心肌细胞 ,进行热应激。用化学发光法测定bcl 2基因转染对热应激心肌细胞线粒体H+ ATPase合成活力的影响 ,用荧光分光光度法测定bcl 2基因转染对热应激心肌细胞Caspase3活性的影响。 结果 :bcl 2基因转染可以使 4 1℃和 4 3℃热应激心肌细胞线粒体H+ ATPase合成活力与转染前相比显著升高 (P <0 .0 1) ,可以使 4 1℃和 4 3℃热应激心肌细胞Caspase3活性与转染前相比显著降低 (P <0 .0 1)。结论 :bcl 2基因转染对热应激心肌细胞凋亡的保护作用可能与其保护心肌细胞线粒体H+ ATPase合成活力 ,并最终阻抑Caspase3活化有关
Objective: To investigate the mechanism of bcl 2 gene transfection on the protection of heat-stressed cardiomyocytes. Methods: The neonatal rat cardiomyocytes were isolated and cultured. The bcl 2 gene was transfected into cardiomyocytes by lipofection and then subjected to heat stress. The effect of bcl 2 gene transfection on the synthesis of mitochondrial H + ATPase in heat-stressed cardiomyocytes was determined by chemiluminescence method. The effect of bcl 2 gene transfection on the activity of Caspase 3 in heat-stressed cardiomyocytes was measured by fluorescence spectrophotometry. Results: The bcl 2 gene transfection could significantly increase the mitochondrial H + ATPase activity of heat-stressed cardiomyocytes at 41 ℃ and 43 ℃, compared with that before transfection (P0.01) The activity of Caspase3 in 4 ℃ heat-stressed cardiomyocytes was significantly lower than that before transfection (P <0.01). CONCLUSION: The protective effect of bcl 2 gene transfection on cardiomyocyte apoptosis may be related to the protection of cardiomyocyte mitochondrial H + ATPase synthesis activity and finally the inhibition of Caspase 3 activation