Sub-Fractions of Red Blood Cells Respond Differently to Shear Exposure Following Superoxide Treatment

Abstract

Red blood cell (RBC) deformability is an essential component of microcirculatory function that appears to be enhanced by physiological shear stress, while being negatively affected by supraphysiological shears and/or free radical exposure. Given that blood contains RBCs with non-uniform physical properties, whether all cells equivalently tolerate mechanical and oxidative stresses remains poorly understood. We thus partitioned blood into old and young RBCs which were exposed to phenazine methosulfate (PMS) that generates intracellular superoxide and/or specific mechanical stress. Measured RBC deformability was lower in old compared to young RBCs. PMS increased total free radicals in both sub-populations, and RBC deformability decreased accordingly. Shear exposure did not affect reactive species in the sub-populations but reduced RBC nitric oxide synthase (NOS) activation and intriguingly increased RBC deformability in old RBCs. The co-application of PMS and shear exposure also improved cellular deformability in older cells previously exposed to reactive oxygen species (ROS), but not in younger cells. Outputs of NO generation appeared dependent on cell age; in general, stressors applied to younger RBCs tended to induce S-nitrosylation of RBC cytoskeletal proteins, while older RBCs tended to reflect markers of nitrosative stress. We thus present novel findings pertaining to the interplay of mechanical stress and redox metabolism in circulating RBC sub-populations.

Influence of 24 h Simulated Altitude on Red Blood Cell Deformability and Hematological Parameters in Patients with Fontan Circulation

“…RBC were separated according to their cell density to yield three distinct fractions which were previously defined as young RBC (less dense, high deformability), old RBC (highest density, lowest deformability), and middle fraction (RBC between young and old RBC) [35][36][37]. Briefly, six Percoll solutions were prepared using a Percoll stock solution (1.130 g/mL; VWR International GmbH, Darmstadt/Germany) and SAH buffer (containing 26.3 g/L BSA, 132 mmol/L NaCl, 4.6 mmol/L and 1 mmol/L HEPES; pH 7.1) as dilution buffer.…”