Endogenous superoxide dismutase activation by oral administration of riboflavin reduces abdominal aortic aneurysm formation in rats Zhenhai Yu, MD, Keisuke.

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Presentation transcript:

Endogenous superoxide dismutase activation by oral administration of riboflavin reduces abdominal aortic aneurysm formation in rats Zhenhai Yu, MD, Keisuke Morimoto, MD, PhD, Jie Yu, MD, Wulan Bao, MD, PhD, Yutaka Okita, MD, PhD, Kenji Okada, MD, PhD Journal of Vascular Surgery Volume 64, Issue 3, Pages 737-745 (September 2016) DOI: 10.1016/j.jvs.2015.03.045 Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 1 Riboflavin or carboxymethyl cellulose (CMC) was given by gastric gavage once daily, starting 3 days before abdominal aortic aneurysm (AAA) preparation. Treated rats (group R) received 25 mg/kg/d riboflavin, and control rats (control) received CMC continuously. 8-OHdG, 8-Hydroxydeoxyguanosine; DHE, dihydroethidium; PCR, polymerase chain reaction; SOD, superoxide dismutase. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 2 A, Aneurysm formation of the abdominal aorta at 28 days is shown in each group. B, Dilatation ratio in each group at the third, seventh, and 28th days. The dilatation ratio (%) was calculated as [dilatation ratio (%) = (maximal aneurysm diameter/normal aortic diameter) × 100]. Data are expressed as the mean ± standard deviation for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .01. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/d); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 3 Histologic findings of abdominal aortic aneurysm (AAA) at day 28 stained with (A) hematoxylin and eosin and (B) elastica van Gieson (EVG) in each group (upper panels: original magnification ×40, bar = 300 μm; lower panels: original magnification ×400, bar = 100 μm). The arrowheads indicate elastin fibers in the aortic wall. C, Quantitative analysis of elastin content. Data are expressed as the mean ± standard deviation (error bars) for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .01. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/d); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 4 Superoxide dismutase (SOD) activity in aortic walls at day 3 was increased with riboflavin treatment. All data are expressed as the mean ± standard deviation (error bars) for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .05. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/d); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 5 A, In situ detection of superoxide by dihydroethidium (DHE) staining at day 7 (original magnification ×400). Mean fluorescence was quantified and expressed as relative values to normal rats. B, Immunostaining of 8-hydroxydeoxyguanosine (8-OHdG)-positive cells in aneurysm walls at day 7 (original magnification ×400). C, Mean fluorescence DHE was quantified and expressed as relative values to normal rats. D, The number of 8-OHdG-positive cells was counted in a high-power field. All data are expressed as the mean ± standard deviation (error bars) for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .01. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/day); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 6 A, Western blot analysis demonstrating the expression of phosphorylated extracellular signal regulated kinase (p-ERK), and total ERK (t-ERK), and β-actin. B, Expression levels of the proteins were measured and expressed as the ratio of the p-ERK and t-ERK of the target protein to that of β-actin. All data are expressed as mean. *P < .05. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/d); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions

Fig 7 A, Intraaortic tumor necrosis factor (TNF)-α, matrix metalloproteinase (MMP)-2, and MMP-9 messenger RNA expression in aneurysm walls at day 7. All data are expressed as the mean ± standard deviation (error bars) for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .01. B, Immunostaining of CD68-positive cells in aneurysm walls at day 7 are shown (original magnification ×400). C, The number of CD68-positive cells was counted in a high-power field. Data are expressed as mean ± standard deviation (error bars) for six rats per group. Statistical analysis was performed using one-way analysis of variance with post hoc test. *P < .01. Control, rats received carboxymethyl cellulose (CMC); group R, rats received riboflavin (25 mg/kg/d); normal, normal native rats. Journal of Vascular Surgery 2016 64, 737-745DOI: (10.1016/j.jvs.2015.03.045) Copyright © 2016 Society for Vascular Surgery Terms and Conditions