Nitric oxide production contributes to Bacillus anthracis edema toxin-associated arterial hypotension and lethality: ex vivo and in vivo studies in the rat

Abstract

The most important aspect of the present study is the finding that the hypotensive and lethal effects of Bacillus anthracis edema toxin are mediated in part via the production of nitric oxide (NO) and that coadministration of an NO synthase inhibitor with edema toxin challenge has a beneficial survival effect .

Abstract

We showed previously that Bacillus anthracis edema toxin (ET), comprised of protective antigen (PA) and edema factor (EF), inhibits phenylephrine (PE)-induced contraction in rat aortic rings and these effects are diminished in endothelial-denuded rings. Therefore, employing rat aortic ring and in vivo models, we tested the hypothesis that nitric oxide (NO) contributes to ET's arterial effects. Compared with rings challenged with PA alone, ET (PA + EF) reduced PE-stimulated maximal contractile force (MCF) and increased the PE concentration producing 50% MCF (EC ₅₀) ( P < 0.0001). Compared with placebo, l-nitro-arginine methyl-ester ( l-NAME), an NO synthase (NOS) inhibitor, reduced ET's effects on MCF and EC ₅₀ in patterns that approached or were significant ( P = 0.06 and 0.03, respectively). In animals challenged with 24-h ET infusions, l-NAME (0.5 or 1.0 mg·kg ⁻¹·h ⁻¹) coadministration increased survival to 17 of 28 animals (60.7%) compared with 4 of 27 (14.8%) given placebo ( P = 0.01). Animals receiving l-NAME but no ET all survived. Compared with PBS challenge, ET increased NO levels at 24 h and l-NAME decreased these increases ( P < 0.0001). ET infusion decreased mean arterial blood pressure (MAP) in placebo and l-NAME-treated animals ( P < 0.0001) but l-NAME reduced decreases in MAP with ET from 9 to 24 h ( P = 0.03 for the time interaction). S-methyl- l-thiocitrulline, a selective neuronal NOS inhibitor, had effects in rings and, at a high dose in vivo models, comparable to l-NAME, whereas N′-[3-(aminomethyl)benzyl]-acetimidamide, a selective inducible NOS inhibitor, did not. NO production contributes to ET's arterial relaxant, hypotensive, and lethal effects in the rat.