ASTRAGALOSIDE IV ATTENUATES HYPOXIA-INDUCED CARDIOMYOCYTE DAMAGE IN RATS BY UPREGULATING SUPEROXIDE DISMUTASE-1 LEVELS
ASTRAGALOSIDE IV ATTENUATES HYPOXIA-INDUCED CARDIOMYOCYTE DAMAGE IN RATS BY UPREGULATING SUPEROXIDE DISMUTASE-1 LEVELS
复制标题
黄芪甲苷 IV 通过上调超氧化物歧化酶 1 水平减轻缺氧引起的大鼠心肌细胞损伤
DOI：
10.1111/j.1440-1681.2008.05059.x
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发表时间：
2009-04-01
影响因子：
2.9
通讯作者：
Huang, Yue-Sheng
中科院分区：
文献类型：
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作者：
Hu, Jiong-Yu;Han, Jian;Huang, Yue-Sheng
1.黄芪甲苷IV（AST-IV）是从天然植物产品中纯化的。先前的研究表明AST-IV具有抗氧化活性。在本研究中，我们研究了AST-IV对大鼠心肌细胞缺氧条件下（长达12 h）的作用和机制.从新生大鼠制备心肌细胞，并在不存在或存在AST-IV（12.5、25或50 μ g/mL）的常氧或缺氧条件下培养。在这些培养条件下研究细胞活力、丙二醛（MDA）水平、超氧化物歧化酶（SOD）-1（mRNA和蛋白质水平分别通过逆转录-聚合酶链反应和蛋白质印迹法测定）和活性氧（ROS;通过2 '，7'-二氯二氢荧光素二乙酸酯测定）的活性和表达。用异硫氰酸荧光素标记的AST-IV检测AST-IV的细胞内定位。缺氧培养降低了心肌细胞的活力，这是改善后，用25或50 μ g/mL的AST-IV治疗。在低氧条件下，MDA水平是对照条件下的两倍。黄芪甲苷IV（25和50 μ g/mL）剂量依赖性地减少缺氧心肌细胞中MDA的增加.异硫氰酸荧光素标记的AST-IV进入心肌细胞并主要定位于细胞质内。5.与常氧相比，低氧条件下，SOD-1活性降低，但mRNA和蛋白表达增加。用25 μ g/mL AST-IV处理后，在常氧和缺氧条件下SOD-1活性和表达均增加。在SOD抑制剂二乙基二硫代氨基甲酸钠（25 μ mol/L）存在下，AST-Ⅳ的ROS清除作用被消除.这些体外结果表明，AST-IV在缺氧条件下保护心肌细胞免受氧化应激介导的损伤。这种作用的主要部分是通过上调细胞质内的SOD-1含量和活性来实现的。
1. Astragaloside IV (AST-IV) is purified from a natural plant product. Previous studies have shown that AST-IV has antioxidant activity. In the present study, we investigated the effect and mechanism of action AST-IV on rat cardiomyocytes subjected to hypoxic conditions (up to 12 h).2. Cardiomyocytes were prepared from neonatal rats and cultured under normoxic or hypoxic conditions in the absence or presence of AST-IV (12.5, 25 or 50 mu g/mL). Cell viability, malondialdehyde (MDA) levels, activity and expression of superoxide dismutase (SOD)-1 (mRNA and protein levels determined by reverse transcription-polymerase chain reaction and western blotting, respectively) and reactive oxygen species (ROS; determined by 2',7'-dichlorodihydrofluorescein diacetate) were investigated under these culture conditions. Intracellular localization of AST-IV was tested using fluorescein isothiocyanate-labelled AST-IV.3. Hypoxic culture reduced the viability of cardiomyocytes, which was improved following treatment with 25 or 50 mu g/mL AST-IV. Under hypoxic conditions, MDA levels were double those under control conditions. Astragaloside IV (25 and 50 mu g/mL) dose-dependently reduced the increase in MDA seen in hypoxic cardiomyocytes.4. Fluorescein isothiocyanate-labelled AST-IV entered cardiomyocytes and was localized mainly within the cytoplasm.5. Under hypoxic conditions, SOD-1 activity was decreased, but mRNA and protein expression increased, compared with normoxia. Following treatment with 25 mu g/mL AST-IV, SOD-1 activity and expression were increased under both normoxic and hypoxic conditions. The ROS scavenging effect of AST-IV was abolished in the presence of the SOD inhibitor sodium diethyl dithiocarbamate (25 mu mol/L).6. These in vitro results show that AST-IV protects cardiomyocytes from oxidative stress-mediated injury under hypoxic conditions. A major part of this action is achieved by upregulation of SOD-1 content and activity within the cell cytoplasm.