吡那地尔、美托洛尔、谷氨酰胺、胰岛素单独及联合使用对H9c2心肌细胞缺氧/复氧损伤的抗氧化作用及机制
首发时间:2011-05-20
摘要:目的:研究吡那地尔(pinacidil, Pin)、美托洛尔(metoprolol, Met)、谷氨酰胺(glutamine, Glu)、胰岛素(insulin, Ins)四药联用对缺氧/复氧(hypoxia/reoxygenation, H/R)所致H9c2心肌细胞损伤的保护作用并探讨其作用机制。方法:将培养的H9c2心肌细胞随机分为7组,即(1) 正常对照(Con)组;(2) 缺氧/复氧(H/R)组;(3) 吡那地尔(Pin)组;(4) 美托洛尔(Met)组;(5) 谷氨酰胺(Glu)组;(6) 胰岛素(Ins)组;(7) 吡那地尔、美托洛尔、谷氨酰胺和胰岛素四药联用(PMGI)组。测定各组H9c2心肌细胞的存活率;收集培养液测定乳酸脱氢酶(lactate dehydrogenase, LDH)活性,超氧化物歧化酶(superoxide dismutase, SOD)活性;Western印迹法检测心肌细胞缺氧/复氧损伤保护性蛋白HSP70的表达情况。结果:对于缺氧/复氧损伤的H9c2心肌细胞,吡那地尔、美托洛尔、谷氨酰胺与胰岛素四药联用组与药物单用组或H/R组相比能明显提高细胞存活率,保护其细胞膜,减少LDH渗漏;增加抗氧化能力,提高SOD含量;增加HSP70的表达。结论:药物联用组与药物单用组相比使心肌细胞抗氧化能力增强,损伤程度减低。
关键词: 缺氧/复氧损伤 乳酸脱氢酶 超氧化物歧化酶 热休克蛋白 抗氧化
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Antioxidation Effects And Mechanism Of Pinacidil, Metoprolol, Glutamine, Insulin Single And Combination On H9c2 Cardiac Myocytes Exposed To Simulated Hypoxia/Reoxygenation
Abstract:Objective:To investigate protective effects and mechanism of pinacidil, metoprolol, glutamine, insulin on H9c2 cardiac myocytes exposed to simulated hypoxia/reoxygenation. Methods:H9c2 cardiac myocytes were randomly divided into seven groups: (1) Control group: cardiomyocytes were seeded in normoxic incubation; (2) Hypoxia/reoxygenation (H/R) group; (3) Pinacidil (Pin) group; (4) Metoprolol (Met) group; (5) Glutamine (Glu) group; (6) Insulin (Ins) group; (7) Pinacidil + Metoprolol + Glutamine + Insulin (PMGI) group. The survival rates of cardiomyocytes were detected. The activity of lactate dehydrogenase (LDH) and superoxide dismutase (SOD) in the culture medium were measured. Western blot was performed to exam expression of heat shock protein 70 (HSP70) in cardiomyocytes respectively. Results: The combination therapy of pinacidil, metoprolol, glutamine, insulin can protect the H9c2 cardiac cell membrane caused by hypoxia/reoxygenation injury; increase cell survival rate, decrease LDH leakage, protect cardiac cell against H/R injury through elevating T-SOD and Mn-SOD activity, increase the expression of HSP70. Conlusion: Compared to the individual drug ,the combination therapy enhances anti-oxidant ability and decrease the injury of the cardiac cell.
Keywords: Hypoxia/Reoxygenation Lactate dehydrogenase Superoxide dismutase Heat shock protein Antioxidation
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吡那地尔、美托洛尔、谷氨酰胺、胰岛素单独及联合使用对H9c2心肌细胞缺氧/复氧损伤的抗氧化作用及机制
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