Nitrite-Mediated Hypoxic Vasodilation Predicted from Mathematical Modeling and Quantified from in Vivo Studies in Rat Mesentery.

Donald G Buerk,Yien Liu,Kelly A Zaccheo,Dov Jaron,Kenneth A Barbee

doi:10.3389/fphys.2017.01053

Abstract

Nitric oxide (NO) generated from nitrite through nitrite reductase activity in red blood cells has been proposed to play a major role in hypoxic vasodilation. However, we have previously predicted from mathematical modeling that much more NO can be derived from tissue nitrite reductase activity than from red blood cell nitrite reductase activity. Evidence in the literature suggests that tissue nitrite reductase activity is associated with xanthine oxidoreductase (XOR) and/or aldehyde oxidoreductase (AOR). We investigated the role of XOR and AOR in nitrite-mediated vasodilation from computer simulations and from in vivo exteriorized rat mesentery experiments. Vasodilation responses to nitrite in the superfusion medium bathing the mesentery equilibrated with 5% O2 (normoxia) or zero O2 (hypoxia) at either normal or acidic pH were quantified. Experiments were also conducted following intraperitoneal (IP) injection of nitrite before and after inhibiting XOR with allopurinol or inhibiting AOR with raloxifene. Computer simulations for NO and O2 transport using reaction parameters reported in the literature were also conducted to predict nitrite-dependent NO production from XOR and AOR activity as a function of nitrite concentration, PO2 and pH. Experimentally, the largest arteriolar responses were found with nitrite >10 mM in the superfusate, but no statistically significant differences were found with hypoxic and acidic conditions in the superfusate. Nitrite-mediated vasodilation with IP nitrite injections was reduced or abolished after inhibiting XOR with allopurinol (p < 0.001). Responses to IP nitrite before and after inhibiting AOR with raloxifene were not as consistent. Our mathematical model predicts that under certain conditions, XOR and AOR nitrite reductase activity in tissue can significantly elevate smooth muscle cell NO and can serve as a compensatory pathway when endothelial NO production is limited by hypoxic conditions. Our theoretical and experimental results provide further evidence for a role of tissue nitrite reductases to contribute additional NO to compensate for reduced NO production by endothelial nitric oxide synthase during hypoxia. Our mathematical model demonstrates that under extreme hypoxic conditions with acidic pH, endogenous nitrite levels alone can be sufficient for a functionally significant increase in NO bioavailability. However, these conditions are difficult to achieve experimentally.

Highlights

The primary source of the nitrite anion (NO−2 ) in mammalian systems is from the oxidation of nitric oxide (NO) produced by the L-arginine/NO enzymatic pathway in vascular endothelium by the O2-dependent endothelial isoform of NO synthase
There were increases in capillary blood flow during NaNO2 superfusion with acidic pH (Figure 1C, solid circles), the difference compared with normal pH was not statistically significant
Our in vivo results from the rat mesentery microcirculation provide further evidence that nitrite reductases in tissue play a role in increasing NO bioavailability during nitrite-mediated hypoxic vasodilation

Summary

Introduction

The primary source of the nitrite anion (NO−2 ) in mammalian systems is from the oxidation of nitric oxide (NO) produced by the L-arginine/NO enzymatic pathway in vascular endothelium by the O2-dependent endothelial isoform of NO synthase (eNOS). Physiological effects of nitrite on the cardiovascular system have been known since 1880 (Reichert and Mitchell, 1880), the consensus view had been that nitrite is an inert byproduct of NO production. Earlier studies using much higher doses of nitrite reported incidences of severe hypotension and lethal methemoglobinemia (Weiss et al, 1937; Wilkins et al, 1937), which curtailed further interest in therapeutic applications for decades. Despite these observed negative effects, nitrite is an approved therapeutic antidote for cyanide and hydrogen sulfide poisoning (Lloyd, 1957; Smith and Gosselin, 1979). Many more dietary studies evaluating the effect of oral nitrate supplements are listed

Methods

Results

Conclusion