Canagliflozin attenuates the progression of atherosclerosis and inflammation process in APOE knockout mice

Figures

• 

  Fig. 1. Serum lipid and fasting blood glucose levels in Cana- and control-groups after 5 weeks of canagliflozin/vehicle oral administration. Significant reduction in total cholesterol, triglyceride, LDL-cholesterol and fasting blood glucose levels was observed in Cana-group at the end of experimental procedure compared to baseline. Fasting glucose was the only significantly increased parameter observed in the control group at the end of intervention. Data are shown as mean ± SD (***P ≤ 0.001, **P ≤ 0.01)

• 

  Fig. 2. Fasting glucose, total cholesterol and triglyceride changes from baseline. Significant changes were detected from baseline in fasting glucose, total cholesterol and triglyceride between study groups. Data are shown as mean ± SD (***P < 0.001, **P < 0.01, *P < 0.05)

• 

  Fig. 3. Canagliflozin/vehicle oral administration effect on heart rate of APOE(−/−) mice. a 5 weeks of canagliflozin intervention led to significant reduction of heart rate (**P < 0.01) while no significant difference was observed in the control group. b Heart rate changes from baseline were significantly different between Cana- and control-groups (*P < 0.05)

• 

  Fig. 4. Atherosclerotic plaque extension among APOE(−/−) mice on western diet treated with canagliflozin (Cana-group) or vehicle (control-group). a Selected 4 μm section images from the aortic root stained with H&E and Masson trichrome. Formation of atherosclerotic plaque was examined using H&E staining while histological examination of atherosclerotic plaque collagen content was assessed using Masson trichrome staining. b Quantification of plaque area is shown as a percentage of lumina stenosis by thickened intima. Collagen content was measured using quantification of Masson trichrome positive area over complete plaque area. Values are shown as mean ± SD and *P < 0.05. Original magnification ×40. H&E indicates hematoxylin and eosin

• 

  Fig. 5. a VCAM-1, MCP-1 and TIMP-1 mRNA expression from thoracic aorta of mice treated with canagliflozin (10 mg/kg/day—5 weeks) and control-group (vehicle—5 weeks). VCAM-1 mRNA expression was significantly decreased in Cana-group as well as MCP-1, while TIMP-1 mRNA expression was increased. b TIMP-1/MMP-2 ratio mRNA was also increased in Cana-group (compared to control-group) approaching the borderline of significance (P = 0.07). Data are presented as mean (**P < 0.01)

• 

  Fig. 6. Atherosclerotic lesion characteristics in APOE(−/−) mice fed with HFD and treated with canagliflozin (Cana-group) or vehicle (control-group). a Selected 4 μm image sections from the aortic root, immunostained for smooth muscle cells (α-Actin), MCP-1, MMP-9 and MMP-2, macrophages (CD68), and TIMP-1/TIMP-2. b Immunostaining differences between groups. Positive cell proportion stained with each antibody was scored from 0 to 4 and staining intensity from 0 to 3 with 0 representing no visual staining and 4 representing intense staining (> 75% of area). Immunoreactivity was scored on a 0–7 scale [extend of staining (0–4) plus intensity of staining (0–3)]. There was a significant difference in MCP-1 expression between Cana-group and control-group (P = 0.048). A-actin and CD68 expression were numerically higher in the Control-group (P = 0.1). Bar graphs show mean ± SD and *P < 0.05. (Original magnification ×200; scale bar, 150 μm)