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目的 研究缺氧对心肌细胞内游离钙离子浓度和肌浆网上的钙离子ATP酶(SERCA2 )的表达的影响及薯蓣皂甙的干预作用。方法 在离体心肌细胞培养基础上制备心肌细胞缺氧模型 ,随机分为 4组 :正常对照组 ,缺氧损伤组 ,薯蓣皂甙组 ,钙拮抗剂组。分别测定各组心肌细胞内游离钙离子的浓度 (FURA2 /AM荧光探针法 )和SERCA2的含量 (Westernblot法 )。结果 缺氧损伤组心肌细胞内游离钙离子的浓度较正常对照组明显增加 ,SERCA2的表达明显减弱 (P均 <0 0 1 ) ;薯蓣皂甙组和钙拮抗剂组心肌细胞内游离钙离子的浓度较缺氧损伤组明显降低 (P <0 0 1 ) ,SERCA2的表达明显增强 (P <0 0 5 )。结论 薯蓣皂甙通过增强肌浆网上钙泵SERCA2的表达减轻缺氧心肌细胞的钙超负荷 ,它对缺氧心肌细胞具有保护作用 ,其作用与钙拮抗剂类似。
Objective To investigate the effects of hypoxia on the intracellular free calcium concentration in myocardium and the expression of calcium ion ATPase (SERCA2) in sarcoplasmic reticulum and the intervention of diosgenin. Methods Myocardial cell hypoxia model was prepared on the basis of cultured in vitro cardiomyocytes, and randomly divided into 4 groups: normal control group, hypoxia injury group, diosgenin group and calcium antagonist group. The concentrations of intracellular free calcium (FURA2 / AM) and SERCA2 in each group were determined by Western blotting. Results The concentration of intracellular free calcium in cardiomyocytes in hypoxia group was significantly higher than that in normal control group, the expression of SERCA2 was significantly decreased (all P <0.01); the concentration of intracellular free calcium in cardiomycin group and calcium antagonist group Compared with hypoxic group, the expression of SERCA2 was significantly decreased (P <0 05). Conclusion Diosgenin can reduce the calcium overload of hypoxic cardiomyocytes by increasing the expression of SERCA2, an on-line calcium pump on the sarcoplasmic reticulum. It has a protective effect on hypoxic cardiomyocytes and its effect is similar to that of calcium antagonists.