去白细胞悬浮红细胞制剂储存过程中活性氧族及丙二醛含量变化

Content changes of reactive oxygen species and malonaldehyde in suspended red blood cells without leukocyte during storage

目的 探讨去白细胞悬浮红细胞制剂储存过程中活性氧族（ROS）含量及丙二醛（MDA）含量的变化.方法 从2013年5～6月于河北省血液中心制备的去白细胞悬浮红细胞制剂中,采用简单随机抽样方法抽取6份去白细胞悬浮红细胞制剂标本作为研究对象.于储存后第0,7,14,28,42天,采用荧光探针2＇,7＇-二氧荧光黄双乙酸盐（DCFH-DA）孵育的方法观察去白细胞悬浮红细胞荧光强度,并对去白细胞悬浮红细胞制剂内ROS含量与MDA含量进行检测.对储存后去白细胞悬浮红细胞制剂内ROS含量、MDA含量分别与储存时间进行相关性分析;并对储存后去白细胞悬浮红细胞制剂内ROS含量与MDA含量进行相关性分析.结果 ①随着储存时间的延长,经荧光探针DCFH-DA孵育的去白细胞悬浮红细胞的荧光强度显著增强;②储存后第0,7,14,28,42天去白细胞悬浮红细胞制剂内ROS含量呈显著增高趋势,第42天去白细胞悬浮红细胞内ROS含量显著高于第0,7,14,28天ROS含量,并且差异均有统计学意义（P＜0.01,0.05,0.05,0.05）;且去白细胞悬浮红细胞制剂内ROS含量与储存时间呈正相关关系（r=0.898,P＜0.01）;③储存后第0,7,14,28,42天去白细胞悬浮红细胞内MDA含量呈显著增高趋势,第42天去白细胞悬浮红细胞内MDA含量显著高于第0,7,14,28天MDA含量,并且差异均有统计学意义（P＜0.01,0.01,0.01,0.05）;且去白细胞悬浮红细胞内MDA含量与储存时间呈正相关关系（r=0.930,P＜0.01）;④储存后第0,7,14,28,42天,去白细胞悬浮红细胞制剂内ROS含量均分别与对应时间点MDA含量呈正相关关系（r=0.877,0.872,0.823,0.786,0.907;P＜0.05）.去白细胞悬浮红细胞制剂储存后第0,7,14,28,42天,各时间点ROS含量均值与MDA含量均值亦呈正相关性关系（r=0.981,P＜0.01）.结论 去白细胞悬浮红细胞制剂储存过程中ROS含量与MDA含量均显著增高,红细胞内氧化应激反应增强,其为导致去白细胞悬浮红细胞制剂储存损伤的重要原因.

Objective To investigate the changes of reactive oxygen species （ROS） concentration and malonaldehyde （MDA） concentration in suspended red blood cells without leukocyte during storage.Methods From May 2013 to June 2013,6 bags of suspended red blood cells without leukocyte which were prepared at Hebei Blood Center were collected in this study by random sampling method.On day 0,day 7,day 14,day 28 and day 42 of storage,the expression of ROS in suspended red blood cells without leukocyte were observed by fluorescent probe 2＇,7＇-dichlorofluorescin diacetate （DCFH-DA） incubation method,the ROS concentration and MDA concentration in suspended red blood cells without leukocyte were also detected.Correlation analysis between the ROS concentration,MDA concentration in suspended red blood cells without leukocyte and storage time was carried out,respectively.Correlation analysis between the MDA concentration and ROS concentration in suspended red blood cells without leukocyte after storage was carried out.Results ① With the extension of storage time,fluorescence intensity of suspended red blood cells without leukocyte incubated with fluorescent probe DCFH-DA was significantly enhanced.② On day 0,day 7,day 14,day 28 and day 42 after storage,ROS concentration in suspended red blood cells without leukocyte showed a significant increasing trend.Concentration of ROS in suspended red blood cells without leukocyte on day 42 was significantly higher than that of day 0,day 7,day 14 and day 28 （P〈0.01,0.05,0.05,0.05）.There was a positive correlation between the ROS concentration in suspended red blood cells without leukocyte and storage time（r=0.898,P〈0.01）.③ On day 0,day 7,day 14,day 28and day 42 after storage,MDA concentration in suspended red blood cells without leukocyte showed a significant increasing trend,MDA concentration in suspended red blood cells without leukocyte on day 42 was significantly higher than that of day 0,day 7,day 14 and day 28 （P〈0.01,0.01,0.01,0.05）.There was a positive correlation between the MDA concentration in suspended red blood cells without leukocyte and storage time （r=0.930,P〈0.01）.④ On day 0,day 7,day 14,day 28 and day 42 after storage,there was a positive correlation between the ROS concentration in suspended red blood cells without leukocyte and MDA concentration of corresponding time respectively （r=0.877,0.872,0.823,0.786,0.907; P〈0.05）.There was also a positive correlation between the mean of ROS concentration of each day and the mean of MDA concentration on day 0,day 7,day 14,day 28 and day 42 after storage （r=0.981,P〈0.01）.Conclusions Concentration of ROS and MDA in suspended red blood cells without leukocyte were significantly increased during storage,and oxidative stress reaction was enhanced in red blood cells.This was the important cause leading to the storage lesions of suspended red blood cells without leukocyte.