Alterations of NO and ATP level in stored RBC incubated with SNP at 37℃
XING Yanfen;WU Qianyu;XIE Xuhong;CUI Yejia;YUAN Zhaohu;YANG Xiuhua;WEI Yaming;Department of Blood Transfusion,Guangzhou First People's Hospital,Guangzhou Medical University;
Objective To investigate the alterations of NO and ATP levels in stored RBC when incubated with NO donor SNP at 37℃. Methods Five blood samples( 100 m L / each) were collected in CPDA-1 from healthy donors and to make suspension RBC. Each blood sample was divided into 4 groups: fresh RBC group( collected within 1 h),stored RBC group( stored for 20 days),added RBC group( 20 days stored RBC incubated with 1,5 and 25 μmol/L SNP at 37℃ for 30min),and SNP addition stored group( incubated RBC with 5 μmol / L SNP for 30 min,and stored RBC to 4℃,then NO and ATP were assayed at 0 h,1 h and 4 h). The total NO,Cytoplasm NO and NO-hemoglobin levels were detected by Total Nitric Oxide assay kit and the ATP level was tested by ATP assay kit. Results Total NO,cytoplasm NO,NO-Hb levels( μmol /L) and ATP levels( μmol / g Hb)( 7. 63 ± 2. 62 vs 15. 56 ± 2. 26,2. 21 ± 1. 68 vs 5. 81 ± 1. 79,5. 42 ± 2. 00 vs 9. 75 ± 2. 73,3. 16 ± 0. 91 vs 8. 10 ± 1. 21,P 0. 05) in stored RBC group were significantly decreased compared to the fresh RBC group.When 1,5 and 25 μmol / L SNP were added into stored RBC and incubated for 30 min,5 μmol / L SNP addition significantly increased total NO,cytoplasm NO,NO-Hb and ATP levels( 14. 15 ± 1. 76,5. 82 ± 3. 38,8. 33 ± 2. 69,6. 16 ± 1. 29) compared with 20 days stored RBC( P 0. 05). 25 μmol / L SNP could increase NO level 5- 10 times more than fresh RBC,but did not show more correction effect on ATP level. When 20 days stored RBC were incubated with 5 μmol / L SNP for 30 min and stored back in 4℃ for 1h and 4h,NO and ATP levels were tested once more. The results showed that both NO and ATP levels at 1h were significantly increased compared to the stored group without SNP addition,and recovered to fresh RBC levels,but at 4h,the three NO levels went down again and did not show improvement compared to stored group,however,ATP level still showed a better recovery than stored group. Conclusion 5 μmol / L SNP incubation for 30 min at 37℃ may be beneficial to improve the quality of stored RBC.