High phosphorus diet induces vascular calcification, a related decrease in bone mass and changes in the aortic gene expression

doi:10.1016/j.bone.2009.09.006

Bone

Volume 46, Issue 1, January 2010, Pages 121-128

https://doi.org/10.1016/j.bone.2009.09.006 Get rights and content

Abstract

In chronic kidney disease, hyperphosphatemia has been associated to vascular calcifications. Moreover, the rate and progression of vascular calcification have been related with the reduction of bone mass and osteoporotic fractures, hereby suggesting a strong link between vascular calcification and bone loss. Our aim was to prospectively study the effects of high phosphorus diet on bone mass, vascular calcification and gene expression profile of the arterial wall.

A rat model of 7/8 nephrectomy fed with normal (0.6%) and moderately high (0.9%) phosphorus diet was used. Biochemical parameters, bone mineral density and vascular calcifications were assessed. A microarray analysis of the aortic tissue was also performed to investigate the gene expression profile.

After 20 weeks, the rats fed with a high phosphorus diet showed a significant increase in serum phosphorus, PTH, and creatinine, together with aortic calcification and a decrease in bone mass. The histological analysis of the vascular calcifications showed areas with calcified tissue and the gene expression profile of this calcified tissue showed repression of muscle-related genes and overexpression of bone-related genes, among them, the secreted frizzled related proteins, well-known inhibitors of the Wnt pathway, involved in bone formation.

The study demonstrated prospectively the inverse and direct relationship between vascular calcification and bone mass. In addition, the microarrays findings provide new information on the molecular mechanisms that may link this relationship.

Introduction

Bone and mineral disorders caused by chronic kidney disease (CKD), particularly by the related poor control of hyperphosphatemia, have been linked to the increased risk of vascular calcification [1], [2]. In addition, the latter has been described as one of the main drivers of mortality in CKD patients [3], [4].

Vascular calcification occurs trough a complex mechanism [5] that consists not only in a simple deposition of calcium and phosphate, but also in a regulated process similar to bone formation which involves loss of vascular calcification inhibitors [6], formation of calcification vesicles [7], and cellular phenotypic changes from vascular smooth muscle cells (VSMCs) to bone-like cells [8]. These new “vascular osteoblasts” are able to express bone-related genes and proteins, such as alkaline phosphatase, Cbfa1/Runx2, osteopontin, osteoprotegerin (OPG) and Msx2, among others [9]. In addition, in the general population and also in CKD patients, the rate and progression of vascular calcification have been associated with an increased risk of bone loss and osteoporotic fractures [10], [11], involving several but not fully understood genetic and molecular mechanisms.

The present experimental study was designed to prospectively assess the effects of high phosphorus diet on bone mass and vascular calcification, while paying attention to the gene expression profile of the arterial wall.

Section snippets

Animal model

The study was performed using 4-month-old male Wistar rats (n = 70). The rats were anaesthetized using methoxi-fluorane, and chronic renal failure (CRF) was induced by surgical 7/8 nephrectomy using the technique modified by Ormrod and Miller [12] by the same technician for all the groups. The nephrectomized rats were subsequently divided in two groups: Group I was fed with normal phosphorus diet (NPD) (0.6% phosphorus, 0.6% calcium and 20% protein content), and group II was fed with high

Biochemical markers

Serum phosphorus and iPTH levels were significantly higher (p < 0.05) in all high phosphorus diet (HPD) groups compared to the paired normal phosphorus diet (NPD) groups. In addition, in the HPD groups, both serum phosphorus and iPTH increased over-time. Conversely, serum calcium was lower in the HPD groups compared to the paired NPD groups, achieving statistically significant differences in the 8HPD and 20HPD groups (Table 1).

All groups showed a reduction in renal function compared to the

Discussion

Vascular calcification, bone loss, increased fractures and high risk of mortality are severe and threatening outcomes in the CKD population at all stages [10], [19], [20], [21]. The importance of all these factors has been recently stressed and also recognized by K-DIGO with the a new nomenclature for all bone mineral disorders associated to CKD [22]. Elevated serum phosphorus has been described as one of the main pathogenetic players for all these abnormalities [2], [23], particularly in the

Acknowledgments

The authors wish to thank Dr. Socorro Braga and Dr. Teresa Fernández-Coto for their assistance in the biochemical analyses, Vanessa Loredo for her help with the TRAP staining in methacrylate sections and Daniel Álvarez-Hernández, Ángeles González-Carcedo and specially Ana Rodríguez-Rebollar for the valuable help with the animals, von Kossa and densitometric analysis. We also thank Marino Santirso for the language review.

Disclosure: This work was supported by Fondo de Investigaciones Sanitarias

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High phosphorus diet induces vascular calcification, a related decrease in bone mass and changes in the aortic gene expression

Abstract

Introduction

Section snippets

Animal model

Biochemical markers

Discussion

Acknowledgments

Am. J. Kidney Dis.

Kidney Int.

Kidney Int.

Kidney Int.

Kidney Int.

Kidney. Int.

Kidney Int.

J. Ren. Nutr.

Kidney Int.

Am. J. Kidney Dis.

Vascular calcifications: pathogenesis, management, and impact on clinical outcomes

J. Am. Soc. Nephrol.

Mechanisms of vascular calcification in chronic kidney disease

J. Am. Soc. Nephrol.

Hyperphosphataemia as a cardiovascular risk factor—how to manage the problem

Nephrol. Dial. Transplant.

Vascular calcification mechanisms

J. Am. Soc. Nephrol.

Human vascular smooth muscle cells undergo vesicle-mediated calcification in response to changes in extracellular calcium and phosphate concentrations: a potential mechanism for accelerated vascular calcification in ESRD

J. Am. Soc. Nephrol.

Molecular mechanisms of vascular calcification: lessons learned from the aorta

Arterioscler. Thromb. Vasc. Biol.

Vascular calcifications, vertebral fractures and mortality in haemodialysis patients

Nephrol. Dial. Transplant.

Progression of vascular calcifications is associated with greater bone loss and increased bone fractures

Osteoporos. Int.

Experimental uremia. Description of a model producing varying degrees of stable uremia

Nephron

Aluminum-induced osteogenesis in osteopenic rats with normal renal function

Calcif. Tissue Int.

Rapamycin retards growth and causes marked alterations in the growth plate of young rats

Pediatr. Nephrol.

Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection

Proc. Natl. Acad. Sci. U. S. A.

Simultaneous changes in the calcium-sensing receptor and the vitamin D receptor under the influence of calcium and calcitriol

Nephrol. Dial. Transplant.

ErmineJ: tool for functional analysis of gene expression data sets

BMC Bioinformatics