Objective To investigate the correlation between the change of adenosine triphosphate(ATP),2,3-diphosphoglycerate(2,3-DPG)concentration and oxidative stress level in suspended red blood cells without leukocyte during storage.Methods The blood of volunteers were collected to prepare suspended red blood cells without leukocyte.The suspended red blood cells were stored at 2-6 ℃.On day 7,day 14,day 28 and day 42 of storage,the concentration of reactive oxygen species(ROS)in red blood cells was detected by fluorescence microplate reader, the concentration of hydrogen peroxide(H2 O2 )outside cell was detected with spectrophotometric method,the concentration of ATP was detected with chemical chromatometry,the 2,3-DPG was detected with Uv test method.Correlation analysis between the ROS,H2 O2 ,ATP,2,3-DPG and storage time were carried out,respectively.Correlation analysis between ATP,2,3-DPG and@ROS,H2 O2 were carried out,respectively.Results With the extension of storage time,the concentration of ROS and H2 O2 were significantly increased,the ROS and H2 O2 content on day 42 of storage were significantly higher than that of day 7,day 14 and day 28.There were positive correlation between ROS and storage time,between H2 O2 and storage time.But with the extension of storage time,the concentration of ATP and 2,3-DPG were significantly decreased. The ATP and 2,3-DPG content on day 42 of storage were significantly lower than that of day 7, day 14 and day 28.There were negative correlation between ATP and storage time,between 2,3-DPG and storage time.There were negative correlation between ATP and ROS,between ATP and H2 O2 ,between 2,3-DPG and ROS,between 2,3-DPG and H2 O2 .Conclusion With the extension of storage time,the concentration of ATP and 2,3-DPG in suspended red blood cells without leukocyte were gradually reduced, which is closely related with the increasing of oxidative stress reaction.%目的 探讨去白悬浮红细胞储存过程中三磷酸腺苷(adenosine triphosphate,ATP)、2,3-二磷酸甘油酸(2,3-diphosphoglycerate,2,3-DPG)含量变化与氧化应激的相关性.方法 采集志愿者血液,制备去白悬浮红细胞,于2~6℃保存.于保存7 d、14 d、28 d、42 d时,应用荧光酶标仪测定去白悬浮红细胞内活性氧族(reactive oxygen species,ROS)含量,采用分光光度法测定细胞外过氧化氢(hydrogen peroxide,H2 O2)含量,采用化学比色法测定ATP含量,采用紫外测试法测定2,3-DPG含量.对去白悬浮红细胞内ROS、ATP、2,3-DPG和细胞外H2 O2含量与储存时间进行相关性分析;对去白悬浮红细胞内ATP、2,3-DPG含量与ROS、H2 O2含量进行相关性分析.结果 随着储存时间的延长,去白悬浮红细胞内ROS含量和细胞外H2 O2含量均明显升高,保存42 d的ROS和H2 O2含量明显高于保存7 d、14 d和28 d红细胞的ROS和H2 O2含量,且ROS和H2 O2含量与储存时间均呈正相关.去白悬浮红细胞内ATP、2,3-DPG含量却明显降低,保存42 d的ATP、2,3-DPG含量明显低于保存7 d、14 d和28 d红细胞的ATP、2,3-DPG含量,并且ATP、2,3-DPG含量与储存时间均呈负相关.去白悬浮红细胞内ATP、2,3-DPG含量与ROS、H2 O2含量也呈负相关.结论 随储存时间延长,去白悬浮红细胞内ATP和2,3-DPG含量逐渐降低,与红细胞内氧化应激反应逐渐增强密切相关.
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