Stratton L P, Rudolph A S, Knoll W K, Bayne S, Farmer M C
Bio/Molecular Engineering Branch, Naval Research Laboratory, Washington, D.C. 20375-5000.
Hemoglobin. 1988;12(4):353-68. doi: 10.3109/03630268808998035.
Preventing the oxidation of hemoglobin in solution is one of the major requirements for the successful production and long-term storage of hemoglobin-based blood substitutes. To this end we have studied the effects of antioxidants on the rate of methemoglobin formation and disappearance in solutions of human and bovine hemoglobin at 4 degrees C and 37 degrees C. Ascorbate and desferal (5 mM) were observed to act as prooxidants, increasing the rate of methemoglobin formation at 37 degrees C. Trehalose, mannitol, glucose, and EDTA (5 mM) had no significant effect. Glutathione and NADH (10 mM) were the most effective antioxidants tested, causing a significant decrease in the rate of methemoglobin formation at 37 degrees C for periods of up to 50 hours. The combination of these antioxidants in bovine hemoglobin at 4 degrees C resulted in the reduction of methemoglobin levels to nearly undetectable levels in approximately 150 hours. In addition, NADH and glutathione were found to reduce methemoglobin levels to 10% over a period of 100 hours in a sample of human hemoglobin that had been stored at 4 degrees C for one year and had 60% methemoglobin. These results suggest that the prevention and reversal of methemoglobin formation during the long-term storage of hemoglobin solutions and hemoglobin-based blood substitutes may now be possible.
防止溶液中的血红蛋白氧化是成功生产和长期储存基于血红蛋白的血液替代品的主要要求之一。为此，我们研究了抗氧化剂对人血红蛋白和牛血红蛋白溶液在4℃和37℃下高铁血红蛋白形成和消失速率的影响。观察到抗坏血酸盐和去铁胺（5 mM）作为促氧化剂，在37℃下增加了高铁血红蛋白的形成速率。海藻糖、甘露醇、葡萄糖和EDTA（5 mM）没有显著影响。谷胱甘肽和NADH（10 mM）是测试的最有效的抗氧化剂，在长达50小时的时间内，在37℃下显著降低了高铁血红蛋白的形成速率。在4℃下，这些抗氧化剂在牛血红蛋白中的组合导致高铁血红蛋白水平在大约150小时内降低到几乎检测不到的水平。此外，在4℃下储存一年且高铁血红蛋白含量为60%的人血红蛋白样本中，发现NADH和谷胱甘肽在100小时内将高铁血红蛋白水平降低到10%。这些结果表明，在血红蛋白溶液和基于血红蛋白的血液替代品的长期储存过程中，防止和逆转高铁血红蛋白的形成现在可能是可行的。
