Volume 83, Issue 5, Pages 835-844 (May 2013) Dietary vitamin K and therapeutic warfarin alter the susceptibility to vascular calcification in experimental chronic kidney disease  Kristin M. McCabe, Sarah L. Booth, Xueyan Fu, Navid Shobeiri, Judith J. Pang, Michael A. Adams, Rachel M. Holden  Kidney International  Volume 83, Issue 5, Pages 835-844 (May 2013) DOI: 10.1038/ki.2012.477 Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 1 Flow diagram of the experimental protocol. BW, body weight. Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 2 Serum creatinine (top), phosphate (middle), and the correlation between serum creatinine and phosphate (bottom; y=0.01x+0.19, P<0.001) in rats treated with no adenine (control) or 0.25% adenine (chronic kidney disease, CKD) after 3, 5, and 7 weeks. Control and CKD rats were treated with either high dietary vitamin K (100mg/kg food), low dietary vitamin K (0.1–0.2mg/kg food), or warfarin (0.08–0.1mg/kg body weight/day). *P<0.05 vs. control, †P<0.05 vs. control and 3 weeks, ‡P<0.05 vs. control, 3 and 5 weeks; n=6–16/group (top and middle) n=53 (bottom); error bars represent s.d. Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 3 Tissue calcium levels (nmol/mg tissue) in the renal artery, carotid artery, abdominal aorta, and thoracic aorta, and the correlation between vessel calcium and phosphate levels in control and chronic kidney disease (CKD) animals. Animals treated with no adenine (control) or 0.25% adenine (CKD) along with high dietary vitamin K (HK; 100mg/kg food), low dietary vitamin K (LK; 0.1–0.2mg/kg food), or warfarin (W; 0.08–0.1mg/kg body weight/day). Line represents the mean, shaded box represents a 95% confidence interval of data from control animals; *P<0.05 versus control, †P<0.05 versus CKD-HK and CKD-LK; n=5–16/group (top four graphs); n=84 (control, bottom left), n=120 (CKD, bottom right, y=0.8x+28.5, P<0.001, r2=0.94, for calcified tissues); vertical dotted line represents three s.d. above control (12nmol/mg tissue). Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 4 Calcification of the aorta, renal, and carotid arteries. (a) Von Kossa–stained blood vessels from animals treated with 0.25% dietary adenine (CKD) or (b) 0% dietary adenine (control) along with high dietary vitamin K (100mg/kg food), low dietary vitamin K (0.1–0.2mg/kg food), or warfarin (0.08–0.1mg/kg BW/day). Positive staining, indicated with arrows, denotes the region of calcification in the carotid, renal, and thoracic aorta of the warfarin- and low vitamin K–treated vessels. Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 5 Systolic blood pressure, diastolic blood pressure, pulse pressure, and pulse wave velocity in animals treated with 0.25% adenine (chronic kidney disease, CKD) along with high dietary vitamin K (HK; 100mg/kg food), low dietary vitamin K (LK; 0.1–0.2mg/kg food), or warfarin (W; 0.08–0.1mg/kg body weight/day).†P<0.05 versus CKD-HK and CKD-LK; n=6–16; line represents mean, and shaded boxes represent a 95% confidence interval of data from control animals. Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions

Figure 6 The effect of dietary vitamin K and warfarin treatment on the level of serum uncarboxylated osteocalcin (GluOC) and carboxylated osteocalcin (GlaOC). Serum GluOC and GlaOC in rats treated with no adenine (a, control) or 0.25% adenine (b, chronic kidney disease (CKD)) for 7 weeks. Control and CKD rats were treated with either high dietary vitamin K (HK, 100mg/kg food), low dietary vitamin K (LK, 0.1–0.2mg/kg food), or warfarin (W, 0.08–0.1mg/kg body weight/day). *P<0.05; n=4–15/group; error bars represent s.d. Kidney International 2013 83, 835-844DOI: (10.1038/ki.2012.477) Copyright © 2013 International Society of Nephrology Terms and Conditions