2018 年第 5 期 第 13 卷

人参皂苷Rb1对乳鼠心肌细胞糖氧化代谢率和糖酵解率的影响

Effect of ginsenoside-Rb1 on glucose oxidative metabolism and glycolysis in neonatal rat cardiomyocytes

作者：黄华廷张薇薇戴胜男孔宏亮

英文作者：

单位：110016沈阳，辽宁省人民医院中国医科大学人民医院心脏中心

英文单位：

关键词：缺氧；心肌细胞；人参皂苷Rb1；糖酵解；腺苷酸活化的蛋白激酶

英文关键词：

• 摘要：

• 目的    探讨人参皂苷Rb1对乳鼠心肌细胞生存能力和糖氧化代谢率、糖酵解率及糖醇解关键酶活性的影响。方法    将乳鼠心肌细胞随机分为对照组、缺氧组和缺氧干预组（即人参皂苷Rb1组、ara A组和人参皂苷Rb1+ara A组），人参皂苷Rb1和ara A［腺苷酸活化的蛋白激酶（AMPK）活性抑制剂］浓度分别为200 μmol/L 和500 μmol/L，干预时间均为8 h。噻唑蓝法检测细胞生存能力，放射免疫法测定糖氧化代谢率和糖酵解率，蛋白质印迹法半定量AMPK活性，酶联免疫吸附试验方法检测己糖激酶、磷酸果糖激酶、乳酸脱氢酶活性。结果    ①缺氧组心肌细胞生存能力、糖氧化代谢率、糖酵解率、AMPK活性均低于对照组［(0.726±0.016)比(0.832±0.016)，(0.232±0.013)比(0.302±0.019),(0.266±0.011)比(0.356±0.010),(0.530±0.020)比(0.620±0.030)］，ara A组［(0.680±0.016)、(0.200±0.012)、(0.210±0.020)、（0.000±0.000）］均低于缺氧组，人参皂苷Rb1组［(0.788±0.023)、(0.282±0.013)、(0.320±0.016)、(1.050±0.040)］均高于缺氧组，人参皂苷Rb1+ara A组［(0.760±0.014)、(0.256±0.005)、(0.296±0.011)、(0.890±0.050)］均低于人参皂苷Rb1组,差异均有统计学意义（均P<0.001）。②缺氧组心肌细胞内己糖激酶活性、磷酸果糖激酶活性和乳酸脱氢酶活性均高于对照组，ara A组均低于缺氧组，人参皂苷Rb1组均高于缺氧组，人参皂苷Rb1+ara A组均低于人参皂苷Rb1组,差异均有统计学意义（均P<0.001）。结论    人参皂苷Rb1可改善缺氧心肌细胞的生存能力，此效应与其“优化”葡萄糖代谢尤其改善心肌细胞的糖酵解有关，所有这些效应均至少部分被AMPK调节。

• 【Abstract】Objective    To investigate the effect of ginsenoside-Rb1(Gs-Rb1) on the survivability, glucose metabolic capacity, glycolysis rate and glycolysis key enzyme activity in ischemic neonatal rat cardiomyocytes. Methods    Neonatal rat cardiomyocytes were cultured in vitro and randomly divided into control group, hypoxia group, Gs-Rb1 group(hypoxia＋Gs-Rb1 200 μmol/L), ara A［adenosine monophosphate activated protein kinase（AMPK） inhibitor］ group(hypoxia＋ara A 500 μmol/L) and Gs-Rb1+ara A group（hypoxia＋Gs-Rb1 200 μmol/L＋ara A 500 μmol/L）. After 8 h culture, cell survivability was tested by MTT; glucose metabolic capacity and glycolysis rate were tested by radioimmunoassay; AMPK activity was tested by western blotting; heterophosphatase(HK), phosphofructokinase(PFK) and lactic dehydrogenase(LDH) activities were tested by enzyme-linked immunosorbent assay. Results    Cell survivability, glucose metabolic capacity, glycolysis rate and AMPK activity in hypoxia group were significantly lower than those in control group［(0.726±0.016)vs(0.832±0.016), (0.232±0.013)vs(0.302±0.019), (0.266±0.011)vs(0.356±0.010), (0.530±0.020)vs(0.620±0.030)］; cell survivability, glucose metabolic capacity, glycolysis rate and AMPK activity in ara A group［(0.680±0.016),(0.200±0.012),(0.210±0.020),（0.000±0.000）］ were significantly lower than those in hypoxia group; cell survivability, glucose metabolic capacity, glycolysis rate and AMPK activity in Gs-Rb1 group［(0.788±0.023),(0.282±0.013),(0.320±0.016),(1.050±0.040)］ were significantly higher than those in hypoxia group; cell survivability, glucose metabolic capacity, glycolysis rate and AMPK activity in Gs-Rb1+ara A group［(0.760±0.014),(0.256±0.005),(0.296±0.011),(0.890±0.050)］ were significantly lower than those in Gs-Rb1 group(all P<0.001). HK activity, PFK activity and LDH activity in hypoxia group were significantly higher than those in control group; HK activity, PFK activity and LDH activity in ara A group were significantly lower than those in hypoxia group; HK activity, PFK activity and LDH activity in Gs-Rb1 group were significantly higher than those in hypoxia group; HK activity, PFK activity and LDH activity in Gs-Rb1+ara A group were significantly lower than those in Gs-Rb1 group(all P<0.001). Conclusion    Gs-Rb1 can improve the survival ability of hypoxic cardiomyocytes, which is related to optimizing glucose metabolism and glycolysis regulated by AMPK.

  【Key words】Hypoxia;Cardiomyocytes;Ginsenoside-Rb1;Glycolysis;Adenosine monophosphate activated protein kinase

  【Fund program】Science and Technology Project of Liaoning Province(2015020282); Science and Technology Project of Shenyang（F15-199-1-06）