Resolvin D1 decreases abdominal aortic aneurysm formation by inhibiting NETosis in a mouse model

Objective: Resolvins have been shown to attenuate inflammation, whereas NETosis, the process of neutrophils releasing neutrophil extracellular traps (NETs), produces increased inflammation. It is hypothesized that treatment of animals with resolvin D1 (RvD1) would reduce abdominal aortic aneurysm (AAA) formation by inhibiting NETosis. Methods: Wild-type 8- to 12-week-old C57BL/6 male mice (n = 47) and apolipoprotein E-deficient (ApoE(-/-)) mice (n = 20) were used in two models to demonstrate the effects of RvD1 on AAA growth. In the topical elastase AAA model, wild-type mice were divided into three groups: a deactivated elastase control group, in which sham surgery was performed using deactivated elastase and mice were intravenously injected with phosphate-buffered saline (PBS) once a day until harvest; an elastase group, in which active elastase was used to induce AAA and mice were injected with PBS daily until harvest; and an RvD1-treated group, in which AAA was induced and mice were injected with RvD1 daily until harvest. In the angiotensin II (Ang II)-induced AAA model, ApoE(-/-) mice were fed a high-fat diet and implanted with osmotic infusion pumps containing Ang II (1000 ng/kg/min). The Ang II model was divided into two groups: an Ang II control group, in which Ang II was delivered and mice were injected with PBS daily until harvest; and an RvD1-treated group, in which Ang II was delivered and mice were injected with RvD1 daily until harvest. On postoperative day 3, day 14, or day 28, aortic and blood samples were collected for Western blot, histology, cytokine array, enzyme-linked immunosorbent assay, and gelatin zymography after aortic diameter measurement. Results: The day 14 RvD1-treated group demonstrated 42% reduced AAA diameter compared with the elastase group (P<.001). On postoperative day 3, the RvD1-treated group showed decreased levels of NETosis markers citrullinated histone H3 (P=.04) and neutrophil elastase (P=.002) compared with the elastase group. Among important cytokines involved in AAA formation, interleukin (IL) 1 beta was downregulated (P=.02) whereas IL-10, a protective cytokine, was upregulated (P=.01) in the RvD1-treated group. Active matrix metalloproteinase 2 also decreased in the RvD1-treated group (P=.03). The RvD1-treated group in the Ang II AAA model, a second model, demonstrated reduced AAA diameter compared with the Ang II control group on day 28 (P<.046). The RvD1-treated group showed decreased levels of citrullinated histone H3 on day 3 (P=.002). Cytokines interferon gamma, IL-1 beta, C-X-C motif chemokine ligand 10, monocyte chemotactic protein 1, and regulated on activation, normal T cell expressed and secreted (RANTES) were all decreased on day 28 (P<.05). Conclusions: RvD1-mediated inhibition of NETosis may represent a future medical treatment for the attenuation of AAA growth. Clinical Relevance: There is currently no successful medical therapy for abdominal aortic aneurysm (AAA). Medical therapies aimed at slowing AAA growth could reduce the occurrence of rupture and the need for future repair. Resolvins are a class of naturally occurring compounds believed to promote resolution by inhibiting NETosis and represent a novel approach to attenuate AAA growth.