This work was supported by NIH Grant HL098032, the Institute for Transfusion Medicine, and the Hemophilia Center of Western Pennsylvania.
Red blood cell endothelial nitric oxide synthase does not modulate red blood cell storage hemolysis
Article first published online: 15 AUG 2012
© 2012 American Association of Blood Banks
Volume 53, Issue 5, pages 981–989, May 2013
How to Cite
Kanias, T., Wang, L., Lippert, A., Kim-Shapiro, D. B. and Gladwin, M. T. (2013), Red blood cell endothelial nitric oxide synthase does not modulate red blood cell storage hemolysis. Transfusion, 53: 981–989. doi: 10.1111/j.1537-2995.2012.03850.x
- Issue published online: 9 MAY 2013
- Article first published online: 15 AUG 2012
- Received for publication March 28, 2012; revision received May 29, 2012, and accepted July 2, 2012.
BACKGROUND: The red blood cell (RBC) endothelial nitric oxide synthase (eNOS) has been shown to regulate intrinsic RBC rheologic properties, such as membrane deformability, suggesting that a functional eNOS could be important in RBC viability and function during storage. This study examines the correlation between RBC eNOS deficiency and the propensity of RBCs to hemolyze under selected stress conditions including prolonged hypothermic storage.
STUDY DESIGN AND METHODS: Fresh or stored RBCs from normal and eNOS knockout (KO) mice or from healthy human volunteers were subjected to selected hemolytic stress conditions including mechanical stress hemolysis, osmotic stress hemolysis, and oxidation stress hemolysis and evaluated during standard storage in CPDA-1 solutions.
RESULTS: Fresh RBCs from normal and eNOS KO mice demonstrated comparable susceptibility to hemolysis triggered by mechanical stress (mechanical fragility index 6.5 ± 0.5 in eNOS KO vs. 6.4 ± 0.4 for controls; n = 8-9), osmotic stress, and oxidative stress. Additionally, RBCs from both mouse groups exhibited similar hemolytic profile at the end of 14-day hypothermic storage, analogous to 42 days of human RBC storage. Storage of human RBCs (28 days in CPDA-1) in the presence of NOS cofactors (l-arginine and tetrahydro-l-biopterin) or inhibitor (N5-[imino(methylamino)methyl]-l-ornithine monoacetate) did not affect cell recovery or hemolytic response to the selected stressors.
CONCLUSION: These studies suggest that RBC eNOS does not modulate susceptibility to hemolysis in response to selected stress conditions or prolonged hypothermic storage. Other strategies to increase nitric oxide (NO) bioactivity after prolonged storage utilizing NOS-independent pathways such as the nitrate–nitrite–NO pathway may prove a more promising approach.