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目的:探讨Presenilin1基因对心肌细胞肌浆网钙容量的影响。方法:本实验以AAV-9为载体构建心肌特异性PS1-shRNA重组腺相关病毒,通过尾静脉注射PS1-shRNA重组腺相关病毒干扰大鼠心肌组织中Presenilin1基因的表达,并利用激光扫描共聚焦显微镜检测并记录心肌细胞肌浆网钙容量及舒张期自发钙释放事件的变化;同时,采用western blot技术分析各组大鼠心肌组织中雷诺丁受体及钙泵表达水平的改变。结果:与对照组相比,PS1-shRNA重组腺相关病毒干预组大鼠心肌细胞肌浆网钙容量显著降低,自发钙释放事件明显增加;雷诺丁受体表达水平略降低,钙泵表达水平无显著性变化。结论:Presenilin1基因表达水平降低会引起心肌细胞钙泄漏,从而造成心肌细胞肌浆网钙容量降低。
Objective: To investigate the effect of Presenilin1 gene on the calcium content of sarcoplasmic reticulum in cardiomyocytes. Methods: AAV-9 was used as a vector to construct cardiac-specific PS1-shRNA recombinant adeno-associated virus. PS1-shRNA recombinant adeno-associated virus was injected into caudal vein to interfere the expression of Presenilin1 gene in rat myocardium. Laser scanning confocal The change of sarcolemmal calcium capacity and spontaneous calcium release during diastole were detected and recorded by microscope. Meanwhile, the changes of the expression of RANOD receptor and calcium pump in myocardial tissue of rats in each group were analyzed by western blot. Results: Compared with the control group, PS1-shRNA recombinant adeno-associated virus intervention significantly decreased the calcium content of sarcoplasmic reticulum and spontaneous calcium release in rats, while the expression of Reynolds receptor decreased slightly. The expression level of calcium pump Significant changes. Conclusion: The decrease of Presenilin1 gene expression causes calcium leakage in cardiac myocytes, resulting in decreased calcium content in sarcoplasmic reticulum of cardiomyocytes.