Fig 1.
Schematic illustration of the experimental design.
For the evaluation of plaque development, inflammation, ROS formation, and endothelial function ApoE-/- mice were used after 8 weeks of intraperitoneal application of NaSCN (200 μg every other day) or vehicle (DMSO). For the assessment of neointima formation wild-type mice were subjected to carotid artery wire injury, treated with NaSCN (200 μg every other day) or vehicle for 14 days, and then analyzed.
Table 1.
Blood pressure, body weight, heart rate and cholesterol upon NaSCN treatment.
Fig 2.
Assessment of atherosclerotic plaque formation, monocyte, and neutrophil granulocyte infiltration in ApoE-/- mice upon NaSCN treatment or DMSO as vehicle.
(A) Quantitative analysis of plaque size as a percentage of the total aortic root vessel wall by oil red staining, n = 5–6. (B) Representative histological images of the aortic root (oil red + hematoxylin staining). (C + D) Histological assessment of monocyte and neutrophil granulocyte infiltration as a percentage of the total vessel wall by immunohistological staining, n = 5. Data are presented as the mean ± SEM., *p ≤ 0.05, ***p ≤ 0.005 vs. vehicle.
Fig 3.
Assessment of IL-6 and IL-10 plasma levels and ROS / chlorotyrosine formation in ApoE-/- mice upon NaSCN treatment or DMSO as vehicle.
(A + B) Plasma IL-6 and IL-10 levels upon NaSCN treatment measured by ELISA, n = 4. (C) Measurement of ROS formation in aortic segments by L-012 chemiluminescence, n = 4. (D) Quantification of HOCl-dependent tissue damage via immunohistological staining of 3-cholortyrosine in the atherosclerotic plaque area, n = 4. (E) Representative images of Chlorotyrosine staining, positive control after incubation with 0.005% HOCl for 1 hour, negative control only with secondary anti-body (Red chlorotyrosine, Blue DAPI). Data are presented as the mean ± SEM., n = 4, *p ≤ 0.05, ***p ≤ 0.005 vs. vehicle.
Fig 4.
Measurement of endothelial function in isolated aortic segments of ApoE−/− mice upon NaSCN treatment in organ chamber experiments.
(A) Definition of the maximal endothelial contraction by incubation with increasing phenylephrine concentrations. (B) Assessment of endothelium-independent vasodilation as a percentage of the maximal contraction with increasing concentrations of nitroglycerin. (C) Assessment of endothelium-dependent vasodilation as a percentage of the maximal contraction with increasing concentrations of carbachol. Data are presented as the mean ± SEM, n = 4–5, **p ≤ 0.01, ***p ≤ 0.005 vs. vehicle.
Fig 5.
Evaluation of neointima formation after carotid-artery wire injury in wild-type mice upon NaSCN treatment.
(A) Quantitative analysis of neointima formation by use of hematoxylin/eosin staining as a percentage of the vessel wall. (B) Representative histological images of the injured carotid artery 14 days post injury. Upper panel with hematoxylin/eosin staining. Lower panel with anti-α-smooth-muscle Actin staining (red) and DAPI (blue). Data are presented as the mean ± SEM, n = 5, ***p ≤ 0.005 vs. vehicle.