论文部分内容阅读
目的研究薯蓣皂苷对培养乳鼠心肌细胞缺氧/复氧损伤的保护作用并探讨其作用机制。方法采用体外培养乳鼠心肌细胞,建立缺氧/复氧损伤模型,以薯蓣皂苷进行干预。心肌细胞随机分为5组:空白对照组(C组);缺氧/复氧组(A/R组);薯蓣皂苷(Dioscin)低、中、高剂量干预组(Dioscin+A/R组)。分别观察各组心肌细胞搏动频率;MTT法测细胞存活率;用Rh123荧光探针标记线粒体,检测荧光强度以反映线粒体膜电位(△Ψm)变化;Fluo-3负载心肌细胞,激光扫描共聚焦显微镜观察细胞内钙离子浓度变化。结果薯蓣皂苷干预组心肌细胞搏动频率、细胞存活率、△Ψm与A/R组比较明显升高;细胞内平均钙离子荧光强度显著低于A/R组。结论从细胞水平初步证实薯蓣皂苷预处理心肌细胞对A/R损伤有一定保护作用,保护机制可能涉及对线粒体膜保护和防止细胞内钙超载等。
Objective To study the protective effect of diosgenin on cultured neonatal cardiomyocytes against hypoxia/reoxygenation injury and to explore its mechanism of action. Methods Rat cardiomyocytes were cultured in vitro and hypoxia/reoxygenation injury models were established. Diosgenin was used to intervene. Cardiomyocytes were randomly divided into 5 groups: blank control group (C group); hypoxia/reoxygenation group (A/R group); Dioscin low, middle and high dose intervention group (Dioscin+A/R group) . The beating frequency of myocardial cells in each group was observed, the survival rate of cells was measured by MTT assay, the mitochondria were labeled with Rh123 fluorescent probe, the fluorescence intensity was measured to reflect the change of mitochondrial membrane potential (ΔΨm), and Fluo-3 loaded cardiomyocytes were scanned by laser scanning confocal microscope. Observe changes in intracellular calcium ion concentration. RESULTS: The beating frequency, cell viability, and △Ψm in the cardiomyocyte-treated group were significantly higher than those in the A/R group. The intracellular calcium fluorescence intensity was significantly lower than that in the A/R group. Conclusion It is preliminarily confirmed that the diosgenin pretreatment of cardiomyocytes has a protective effect on A/R injury from the cellular level, and the protection mechanism may involve the protection of mitochondrial membrane and the prevention of intracellular calcium overload.