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目的:探讨人参皂苷Rg1(ginsenoside Rg1)对异丙肾上腺素(Isoprenaline,ISO)诱导大鼠心肌细胞肥大中能量代谢的作用及机制。方法:利用体外培养模型,用异丙肾上腺素(10μmol/L)诱导心肌细胞肥大,观察β受体阻滞剂普萘洛尔(2μmol/L)及不同计量人参皂苷Rg1(12.5、25、50μmol/L)对异丙肾上腺素诱导大鼠心肌细胞肥大的作用。用消化分离法及计算机图像分析系统测细胞体积;细胞荧光染色观察细胞形态大小;考马斯亮蓝(Bradford)法测心肌细胞总蛋白含量;酶联免疫吸附(ELISA)法检测细胞外液中游离脂肪酸(FFA)、单磷酸腺苷(AMP)、二磷酸腺苷(ADP)、三磷酸腺苷(ATP)的含量;反转录-PCR(RT-PCR)法检测心房钠尿肽(ANP)mRNA的表达;蛋白质印迹(Western blot)法检测心肌细胞过氧化物酶体增殖活化受体γ辅助活化因子1α(PGC-1α)、丙酮酸脱氢酶激酶4(PDK4)蛋白的表达水平。结果:与空白对照组相比,异丙肾上腺素模型组心肌细胞体积明显增大,蛋白含量增加,ANP mRNA表达增加,PGC-1α、PDK4蛋白表达减少,心肌组织游离脂肪酸(FFA)升高,ATP/ADP、ATP/AMP明显降低。与异丙肾上腺素组相比,人参皂苷Rg1(12.5、25、50μmol/L)均能够有效的改善异丙肾上腺素诱导的心肌细胞肥大,蛋白含量降低,ANP mRNA表达降低,PGC-1α、PDK4蛋白表达增高,细胞外液中FFA降低,ATP/ADP、ATP/AMP增加,且呈一定的剂量依赖性。结论:人参皂苷Rg1能改善异丙肾上腺素诱导的心肌细胞肥大,其机制可能是调控PGC-1α来提高异丙肾上腺素诱导的心肌细胞肥大能量代谢水平。
AIM: To investigate the effect and mechanism of ginsenoside Rg1 on energy metabolism in rat cardiomyocyte hypertrophy induced by isoprenaline (ISO). Methods: Myocardial cell hypertrophy was induced by isoproterenol (10 μmol / L) in vitro. Propranolol (2 μmol / L) and different concentrations of ginsenoside Rg1 (12.5, 25 and 50 μmol / L) on isoproterenol-induced cardiomyocyte hypertrophy in rats. Cell volume was measured by digestion and computerized image analysis system; Cell morphology was observed by fluorescence staining; Total protein content of myocardial cells was measured by Bradford method; Free fatty acids in extracellular fluid were detected by enzyme-linked immunosorbent assay (ELISA) (FFA), adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). The atrial natriuretic peptide (ANP) mRNA expression was detected by RT- Western blot was used to detect the expression of peroxisome proliferator-activated receptor γ-helper activating factor 1α (PGC-1α) and pyruvate dehydrogenase kinase 4 (PDK4) in cardiomyocytes. Results Compared with the blank control group, the isoproterenol significantly increased the volume of cardiomyocytes, increased the protein content, increased the expression of ANP mRNA, decreased the expression of PGC-1α and PDK4, and increased the level of free fatty acids (FFA) ATP / ADP, ATP / AMP decreased significantly. Compared with isoproterenol group, ginsenoside Rg1 (12.5,25 and 50μmol / L) could effectively improve isoprenaline-induced cardiomyocyte hypertrophy, reduce protein content and decrease the expression of ANP mRNA, while PGC-1α and PDK4 Increased protein expression, decreased FFA in extracellular fluid, ATP / ADP, ATP / AMP increased, and in a dose-dependent manner. CONCLUSION: Ginsenoside Rg1 can improve isoprenaline-induced cardiomyocyte hypertrophy. Its mechanism may be that PGC-1α may be involved in the regulation of isoproterenol-induced hypertrophy and energy metabolism in cardiomyocytes.