Figure 1.
Pharmacokinetics and distribution of ELK-2A2K2E in mouse plasma.
A – Pharmacokinetics of ELK-2A2K2E in Apoe−/− mice fed high- fat diet (representative curve from one animal). B – FPLC analysis of distribution of ELK-2A2K2E and cholesterol in plasma collected 2 h after peptide injection.
Figure 2.
Effect of ELK-2A2K2E treatment on development of atherosclerosis: en face analysis after staining with Sudan IV.
A - Percentage of atherosclerotic lesions in the total aorta; n = 8 for each bar. B - Percentage of atherosclerotic lesions in the aortic arch; *p<0.03; **p<0.05 versus vehicle; n = 8 for each bar. C- Percentage of atherosclerotic lesions in the thoracic aorta; n = 8 for each bar. D - Percentage of atherosclerotic lesions in the abdominal aorta; n = 8 for each bar. Percentages were calculated as an area stained with Sudan IV divided by total area.
Figure 3.
Effect of ELK-2A2K2E treatment on development of atherosclerosis: histological analysis.
A- Quantitation of the area occupied by lesions within the aortic sinus region after staining with Oil Red O; *p<0.02 versus vehicle; n = 8 for each bar. B - Representative section of an aortic sinus from a control mouse stained with Oil Red O. C - Representative section of an aortic sinus from a mouse treated with ELK-2A2K2E stained with Oil Red O. D - Analysis of the abundance of collagen within the aortic sinus region after staining with Masson’s trichrome stain; n = 8 for each bar. Values represent collagen positive staining as a percentage of lesion area. E - Representative section of an aortic sinus from a control mouse stained with Masson’s trichrome stain. F - Representative section of an aortic sinus from a mouse treated with ELK-2A2K2E stained with Masson’s trichrome stain.
Figure 4.
Effect of ELK-2A2K2E treatment on inflammation and oxidation in aorta.
A - Analysis of macrophage infiltration within the aortic sinus region after staining with anti CD68; *p<0.001 versus vehicle; n = 8 for each bar. B - Representative section of an aortic sinus from a control mouse stained with anti CD68. C - Representative section of an aortic sinus from a mouse treated with ELK-2A2K2E stained with anti CD68. D - Analysis of the abundance of VCAM-1 in sections from the aortic sinus region; *p<0.001 versus vehicle; n = 8 for each bar. E - Representative section of an aortic sinus from a control mouse stained with anti VCAM-1. F - Representative section of an aortic sinus from a mouse treated with ELK-2A2K2E stained with anti VCAM-1. G - Analysis of the abundance of nitrotyrosine (NT) in sections from the aortic sinus region; *p<0.001 versus vehicle; n = 8 for each bar. H - Representative section of an aortic sinus from a control mouse stained with anti- nitrotyrosine. I - Representative section of an aortic sinus from a mouse treated with ELK-2A2K2E stained with anti-nitrotyrosine. Values represent positive staining with the corresponding antibody as a percentage of lesion area.
Figure 5.
Effect of ELK-2A2K2E on plasma lipids and lipoproteins.
A - Total cholesterol; B - non-HDL cholesterol; C - Triglycerides; D - HDL cholesterol; E - apoA-I. *p<0.05, **p<0.01 versus vehicle; n = 8 for each point.
Figure 6.
Effect of ELK-2A2K2E on HDL size distribution and HDL functionality.
A - Analysis of size distribution of HDL particles in two groups of mice assessed by non-denaturing electrophoresis; *p<0.02 versus vehicle. B - Cholesterol efflux to plasma (final concentration 1%) from two groups of mice; plasma collected 2 h after peptide injection. C - Cholesterol efflux to plasma (final concentration 1%) from two groups of mice; plasma collected 72 h after peptide injection. D - Efflux of cholesterol to plasma (final concentration 1%), ELK-2A2K2E (final concentration 10 µg/ml) or a combination of the two. *p<0.05 versus plasma.
Figure 7.
Effect of ELK-2A2K2E on the rate of reverse cholesterol transport in vivo.
RAW 264.7 macrophages were labeled with [3H]cholesterol, loaded with AcLDL and transplanted into the intra-peritoneal cavity of C57Bl/6 mice. Proportion of [3H]cholesterol out of total injected in macrophages recovered after 24 h in plasma (A), liver (B), fecal cholesterol (C) and fecal bile acids (D) are shown; *p<0.05; **p<0.001 versus vehicle; #p<0.01 versus 5A; n = 7 for each bar.