Ek B, Hallberg C, Sjögren K G, Hjalmarson A
ASTRA HASSLE AB, Mölndal, Sweden.
Free Radic Biol Med. 1994 Jan;16(1):117-21. doi: 10.1016/0891-5849(94)90248-8.
To study the role of chain-breaking antioxidants on reperfusion injury in the ischemic heart, cultured ventricular heart cells (myocytes) were subjected to hypoxia and reoxygenation. The myocytes were prepared from neonatal rats and cultured in F10 medium that was supplemented with serum. As a marker for cell damage, lactate dehydrogenase was analyzed in the medium. Cells subjected to hypoxia for 5 h showed a 1.9 fold increase in lactate dehydrogenase (LD) leakage, while cells subjected to 1 h hypoxia followed by 4 h reoxygenation showed a 5-fold increase in LD intake. Alpha-tocopherol, beta-carotene, nordihydroguairetic acid (NDGA), butylated hydroxyltoluene (BHT), and ICI211965 were added to the cell medium every 24 h for 6 d prior to reoxygenation. All compounds protected against reoxygenation-induced cell damage. In the presence of the 5-lipoxygenase inhibitor ICI211965, protection against LD leakage was found only at high concentrations, which corresponded to the antioxidative effect of ICI211965, and not to inhibition of 5-lipoxygenase. We conclude that cultured ventricular myocytes can be used to evaluate the protective effect of antioxidants on reoxygenation-induced cell damage, and that chain-breaking antioxidants protected well against reoxygenation-induced cell damage.
为研究断链抗氧化剂在缺血性心脏再灌注损伤中的作用，对培养的心室心肌细胞（心肌细胞）进行缺氧和复氧处理。心肌细胞取自新生大鼠，在添加血清的F10培养基中培养。作为细胞损伤的标志物，对培养基中的乳酸脱氢酶进行分析。缺氧5小时的细胞乳酸脱氢酶（LD）泄漏增加了1.9倍，而缺氧1小时后再复氧4小时的细胞LD摄取增加了5倍。在复氧前6天，每24小时向细胞培养基中添加一次α-生育酚、β-胡萝卜素、去甲二氢愈创木酸（NDGA）、丁基羟基甲苯（BHT）和ICI211965。所有化合物均能保护细胞免受复氧诱导的损伤。在5-脂氧合酶抑制剂ICI211965存在的情况下，仅在高浓度时发现对LD泄漏有保护作用，这与ICI211965的抗氧化作用相对应，而非对5-脂氧合酶的抑制作用。我们得出结论，培养的心室肌细胞可用于评估抗氧化剂对复氧诱导的细胞损伤的保护作用，并且断链抗氧化剂能很好地保护细胞免受复氧诱导的损伤。
