P0919CALORIC RESTRICTION DOES NOT PROTECT AGAINST THE DEVELOPMENT OF VASCULAR CALCIFICATION
Authors: Ríos-Varo Rafael , Vidal Ángela , Raya Ana Isabel , Pineda Carmen , López Ignacio , Rodriguez-Portillo J Mariano , Aguilera-Tejero Escolástico 
Published: June 2020

Abstract

Background and Aims
Vascular calcification (VC) is an important contributor to the high rate of cardiovascular mortality associated to chronic kidney disease.
The inability to eliminate phosphorus (P) and the subsequent P retention promotes CV. P metabolism and uremic VC are influenced by obesity and by the caloric content of the diet.
Caloric restriction (CR) has been shown to have multiple beneficial effects on health, for example, CR has been reported to improve vascular health and retard vascular ageing. However, to our knowledge the effect of CR on the development of uremic VC has not been explored.
We hypothesize that CR may be beneficial to prevent the development of uremic VCs. Thus, the objective of the present study was to determine if rats subjected to CR were protected against VC.

Method
48 Wistar rats were divided in four groups. The control diet provided Metabolizable Energy = 3.528 kcal/g and contained 0.6% Calcium (Ca) and 0.6% P. Additional diets of identical composition to the control diet but containing varying levels of Ca and P: 0.9% Ca, 0.9% P; 0.6% Ca, 1.2% P; and 0.9% Ca, 1.8% P, were also used in the experiments.
Rats in Group 1 and 3 were fed 15 g/day of the control diet. Rats in Group 2 and 4 were calorie restricted and fed 10 g/day of diet with Ca/P = 0.9%/0.9%. Thus the daily P intake should be identical in the four groups.
Uremia was induced by 5/6 nephrectomy (Nx). After Nx rats in Group 1 and 2 were fed ad libitum a diet with 0.6% Ca and 1.2% P. While rats in Group 3 and 4 were fed ad libitum a diet with 0.9% Ca and 1.8% P. Rats were supplemented with calcitriol.
At the end of the experiment, rats were sacrificed to obtain blood samples and tissue samples (thoracic and abdominal aortas).
After blood collection, plasma was separated by centrifugation and stored at –20° C until assayed. Plasma creatinine, Ca and P were measured by spectrophotometry. Energy intake was calculated based on food intake. VC was studied by histology and by measuring the tissue Ca content.
Values are expressed as mean ± standard error (SE), the difference between groups was assessed by ANOVA. Fisher LSD test was used as a post-hoc procedure. p&lt;0.05 was considered significant.

Results
Before Nx, caloric intake was significantly lower in calorie restricted rats (35.4 ± 0.1 and 35.8 ± 0.1 kcal/day) than in rats eating normal calories (52.7 ± 0.1 and 52.8 ± 0.2 kcal/day); however, P intake was almost identical in the four groups and ranged between 89.8 and 91.6 mg/day.
After Nx, rats in all groups reduced food intake and, consequently, caloric intake. Thus, although the P content of the diet was increased after Nx, daily P intake was not increased in Groups 1 and 2; however, P intake was significantly increased in Groups 3 and 4 (120.9 ± 4.6 and 122.2 ± 6.2 mg/day, respectively).
In all groups, rats had high plasma concentrations of creatinine and P, and low plasma concentrations of Ca. Also, all rats had elevated Ca content in the aorta. No significant differences between the study groups were found in any of these parameters (Table 1).
Von Kossa staining of the aortas showed abundant mineral deposition in the four groups.

Conclusion
This study shows that, contrary to what was expected, CR did not prevent or ameliorate uremic calcifications.