Erythrocytic bioactivation of nitrite and its potentiation by far-red light

Background Nitrite is reduced by heme-proteins and molybdenum-containing enzymes to form the important signaling molecule nitric oxide (NO), mediating NO signaling. Substantial evidence suggests that deoxygenated hemoglobin within red blood cells (RBCs) is the main erythrocytic protein responsible for mediating nitrite-dependent NO signaling. In other work, infrared and far red light have been shown to have therapeutic potential that some attribute to production of NO. Here we explore whether a combination of nitrite and far red light treatment has an additive effect in NO-dependent processes, and whether this effect is mediated by RBCs. Methods and results Using photoacoustic imaging in a rat model as a function of varying inspired oxygen, we found that far red light (660 nm, five min. exposure) and nitrite feeding (three weeks in drinking water at 100 mg/L) each separately increased tissue oxygenation and vessel diameter, and the combined treatment was additive. We also employed inhibition of human platelet activation measured by flow cytometry to assess RBC-dependent nitrite bioactivation and found that far red light dramatically potentiates platelet inhibition by nitrite. Blocking RBC-surface thiols abrogated these effects of nitrite and far-red light. RBC-dependent production of NO was also shown to be enhanced by far red light using a chemiluminescence-based nitric oxide analyzer. In addition, RBC-dependent bioactivation of nitrite led to prolonged lag times for clotting in platelet poor plasma that was enhanced by exposure to far red light. Conclusions Our results suggest that nitrite leads to the formation of a photolabile RBC surface thiol-bound species such as an S-nitrosothiol or heme-nitrosyl (NO-bound heme) for which far red light enhances NO signaling. These findings expand our understanding of RBC-mediated NO production from nitrite. This pathway of NO production may have therapeutic potential in several applications including thrombosis, and, thus, warrants further study.
Graphical abstract Potential mechanism of RBC-mediated nitrite bioactivation and its potentiation by far red light. Nitrite reacts with deoxygenated Hb to make NO which then binds other vacant hemes or forms a nitrosothiol (RSNO). The nitrosyl-heme or nitrosothiol is exported and binds a surface thiol. Another potential NO species that may form is a DNIC (not shown). The NO congener can then be transported in plasma and interact with platelets and other blood components and this action is potentiated by photolysis using far red light.fx1
Highlights • Nitrite and far red light increase tissue oxygenation under hypoxia. • Far red light enhances nitrite-mediated inhibition of platelet activation by red cells. • Blocking red cell surface thiols abrogates nitrite-mediated effects. • NO production from nitrite and red cells is enhanced by far red light. • Lag time in clotting is prolonged by nitrite plus far red light.