去白悬浮红细胞储存过程中抗氧化酶活性的改变

Changes of antioxidant enzyme activities in suspended red blood cell without leukocyte during storage

目的:观察去白悬浮红细胞储存期间超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)活性的改变及其与氧化应激的相关性。方法:从河北省血液中心制备的去白悬浮红细胞中随机抽取6袋作为研究对象,置2~6℃冰箱保存。储存第7、14、28、42天使用荧光酶标仪测定红细胞内活性氧族(ROS)含量,硫代巴比妥酸比色法测定细胞内丙二醛(MDA)含量,黄嘌呤氧化酶法测定SOD活性;可见光法测定CAT活性;比色法测定GSH-Px活性。结果:去白悬浮红细胞储存第42天,ROS含量和MDA含量均显著高于储存第7天、14天和28天(P<0.05);细胞内SOD、CAT、GSH-Px活性均显著低于储存第7、14和28天(P<0.05);去白悬浮红细胞内ROS含量、MDA含量与储存时间呈正相关(P<0.05),而SOD活性、CAT活性、GSH-Px活性与储存时间呈负相关(P<0.05);去白悬浮红细胞内SOD活性、CAT活性、GSH-Px活性与细胞内ROS含量、MDA含量呈负相关(P<0.05)。结论:随着储存时间延长,去白悬浮红细胞内抗氧化酶SOD、CAT、GSH-Px活性降低是导致细胞内氧化应激水平和过氧化损伤程度升高的原因之一。

Objective: To observe the change of superoxide dismutase(SOD), catalase(CAT), glutathione peroxidase(GSH-Px) activity in suspended red blood cell without leukocyte during storage and the correlation between activity and oxidative stress. Methods: Six bags of suspended red blood cells without leukocyte prepared in Hebei Blood Center were randomly selected and regarded as research object. The red blood cells were stored in refrigerator at 2-6℃. On 7, 14, 28 and 42 days of storage, the reactive oxygen species(ROS) content in red blood cells was detected with fluorescence microplate reader. The malondialdehyde(MDA) content was detected by TBA method. The activity of SOD was detected by xanthine oxidase method, the activity of CAT was detected by visible light method, and the activity of GSH-Px was detected by colorimetry. Results: On 42 days of storage of suspended red blood cell without leukocyte, the content of ROS and MDA were significantly higher than those of day 7, day 14 and day 28(P<0.05). The activities of SOD, CAT and GSH-Px in cell were significantly lower than those of day 7, day 14 and day 28(P<0.05). There were positive correlation between the ROS content, MDA content and storage time. There were negative correlation between SOD activity, CAT activity, GSH-Px activity and storage time. There were negative correlation between SOD activity, CAT activity, GSH-Px activity and ROS content, MDA content in suspended red blood cells without leukocyte. Conclusion: With the increase of storage time, the decrease of SOD, CAT and GSH-Px activities in suspended red blood cells without leukocyte is one of the reasons for increase of oxidative stress and peroxidation damage degree in red cell.