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Impaired Vitamin D Signaling in Endothelial Cell Leads to an Enhanced Leukocyte-Endothelium Interplay: Implications for Atherosclerosis Development.

Bozic M, Álvarez Á, de Pablo C, Sanchez-Niño MD, Ortiz A, Dolcet X, Encinas M, Fernandez E, Valdivielso JM - PLoS ONE (2015)

Bottom Line: Inhibition of NF-κB activation with super-repressor IκBα blunted all signs of endothelial cell activation caused by downregulation of VDR in endothelial cells.In vivo, deletion of VDR led to significantly larger aortic arch and aortic root lesions in apoE-/- mice, with higher macrophage content. apoE-/-VDR-/-mice showed higher aortic expression of VCAM-1, ICAM-1 and IL-6 when compared to apoE-/-VDR+/+ mice.Our data demonstrate that lack of VDR signaling in endothelial cells leads to a state of endothelial activation with increased leukocyte-endothelial cell interactions that may contribute to the more severe plaque accumulation observed in apoE-/-VDR-/- mice.

View Article: PubMed Central - PubMed

Affiliation: Nephrology Research Department, IRB Lleida, Lleida, Spain.

ABSTRACT
Endothelial cell activation leading to leukocyte recruitment and adhesion plays an essential role in the initiation and progression of atherosclerosis. Vitamin D has cardioprotective actions, while its deficiency is a risk factor for the progression of cardiovascular damage. Our aim was to assess the role of basal levels of vitamin D receptor (VDR) on the early leukocyte recruitment and related endothelial cell-adhesion-molecule expression, as essential prerequisites for the onset of atherosclerosis. Knockdown of VDR in endothelial cells (shVDR) led to endothelial cell activation, characterized by upregulation of VCAM-1, ICAM-1 and IL-6, decreased peripheral blood mononuclear cell (PBMC) rolling velocity and increased PBMC rolling flux and adhesion to the endothelium. shVDR cells showed decreased IκBα levels and accumulation of p65 in the nucleus compared to shRNA controls. Inhibition of NF-κB activation with super-repressor IκBα blunted all signs of endothelial cell activation caused by downregulation of VDR in endothelial cells. In vivo, deletion of VDR led to significantly larger aortic arch and aortic root lesions in apoE-/- mice, with higher macrophage content. apoE-/-VDR-/-mice showed higher aortic expression of VCAM-1, ICAM-1 and IL-6 when compared to apoE-/-VDR+/+ mice. Our data demonstrate that lack of VDR signaling in endothelial cells leads to a state of endothelial activation with increased leukocyte-endothelial cell interactions that may contribute to the more severe plaque accumulation observed in apoE-/-VDR-/- mice. The results reveal an important role for basal levels of endothelial VDR in limiting endothelial cell inflammation and atherosclerosis.

No MeSH data available.


Related in: MedlinePlus

VDR deletion increases aortic and systemic inflammatory responses.(A) Effects of VDR deletion on proinflammatory cytokine production. Circulating serum levels of interleukin (IL-6) were determined by ELISA. Data presented are mean ± SEM of n = 10 mice/group. *p<0.05 vs. apoE-/-. (B, C, D, E, F) Real time PCR analysis of VCAM-1, ICAM-1 and IL-6 mRNA in the aortic tissue of apoE-/- and apoE-/-VDR-/- mice (B, C, D) and VDR+/+ and VDR-/- mice (E, F). Data presented are mean ± SEM. (B, C) *p<0.05 vs. apoE-/-; (E, F) *p<0.05 vs. VDR+/+ mice. (G) Enhanced VCAM-1 and ICAM-1 expression in the vascular endothelium of apoE-/-VDR-/- mice. Aortic root lesions from apoE-/- and apoE-/-VDR-/- mice were stained with anti-VCAM-1 and anti-ICAM-1. CD31 was used for endothelial cell staining. Representative images are shown in G. Magnification 20x.
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pone.0136863.g006: VDR deletion increases aortic and systemic inflammatory responses.(A) Effects of VDR deletion on proinflammatory cytokine production. Circulating serum levels of interleukin (IL-6) were determined by ELISA. Data presented are mean ± SEM of n = 10 mice/group. *p<0.05 vs. apoE-/-. (B, C, D, E, F) Real time PCR analysis of VCAM-1, ICAM-1 and IL-6 mRNA in the aortic tissue of apoE-/- and apoE-/-VDR-/- mice (B, C, D) and VDR+/+ and VDR-/- mice (E, F). Data presented are mean ± SEM. (B, C) *p<0.05 vs. apoE-/-; (E, F) *p<0.05 vs. VDR+/+ mice. (G) Enhanced VCAM-1 and ICAM-1 expression in the vascular endothelium of apoE-/-VDR-/- mice. Aortic root lesions from apoE-/- and apoE-/-VDR-/- mice were stained with anti-VCAM-1 and anti-ICAM-1. CD31 was used for endothelial cell staining. Representative images are shown in G. Magnification 20x.

Mentions: After eight weeks on a HFRD diet, serum IL-6 levels were higher in apoE-/-VDR-/- mice than in apoE-/- control animals (Fig 6A). Furthermore, DKO mice exhibited markedly elevated expression of VCAM-1 (Fig 6B) and ICAM-1 (Fig 6C) in the aortae, as well as higher expression of IL-6 than apoE-/- counterparts (Fig 6D). Immunohistochemical staining of the aortic root lesions showed increased expression of VCAM-1 protein, as well as a mild immunoreactivity for ICAM-1 in the vascular endothelium of DKO mice compared to apoE-/- animals (Fig 6G). In the absence of the apoE- genotype and evident lack of macrophage plaque accumulation, VDR-/- mice still exhibited an increased arterial expression of adhesion molecules (Fig 6E and 6F).


Impaired Vitamin D Signaling in Endothelial Cell Leads to an Enhanced Leukocyte-Endothelium Interplay: Implications for Atherosclerosis Development.

Bozic M, Álvarez Á, de Pablo C, Sanchez-Niño MD, Ortiz A, Dolcet X, Encinas M, Fernandez E, Valdivielso JM - PLoS ONE (2015)

VDR deletion increases aortic and systemic inflammatory responses.(A) Effects of VDR deletion on proinflammatory cytokine production. Circulating serum levels of interleukin (IL-6) were determined by ELISA. Data presented are mean ± SEM of n = 10 mice/group. *p<0.05 vs. apoE-/-. (B, C, D, E, F) Real time PCR analysis of VCAM-1, ICAM-1 and IL-6 mRNA in the aortic tissue of apoE-/- and apoE-/-VDR-/- mice (B, C, D) and VDR+/+ and VDR-/- mice (E, F). Data presented are mean ± SEM. (B, C) *p<0.05 vs. apoE-/-; (E, F) *p<0.05 vs. VDR+/+ mice. (G) Enhanced VCAM-1 and ICAM-1 expression in the vascular endothelium of apoE-/-VDR-/- mice. Aortic root lesions from apoE-/- and apoE-/-VDR-/- mice were stained with anti-VCAM-1 and anti-ICAM-1. CD31 was used for endothelial cell staining. Representative images are shown in G. Magnification 20x.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4556440&req=5

pone.0136863.g006: VDR deletion increases aortic and systemic inflammatory responses.(A) Effects of VDR deletion on proinflammatory cytokine production. Circulating serum levels of interleukin (IL-6) were determined by ELISA. Data presented are mean ± SEM of n = 10 mice/group. *p<0.05 vs. apoE-/-. (B, C, D, E, F) Real time PCR analysis of VCAM-1, ICAM-1 and IL-6 mRNA in the aortic tissue of apoE-/- and apoE-/-VDR-/- mice (B, C, D) and VDR+/+ and VDR-/- mice (E, F). Data presented are mean ± SEM. (B, C) *p<0.05 vs. apoE-/-; (E, F) *p<0.05 vs. VDR+/+ mice. (G) Enhanced VCAM-1 and ICAM-1 expression in the vascular endothelium of apoE-/-VDR-/- mice. Aortic root lesions from apoE-/- and apoE-/-VDR-/- mice were stained with anti-VCAM-1 and anti-ICAM-1. CD31 was used for endothelial cell staining. Representative images are shown in G. Magnification 20x.
Mentions: After eight weeks on a HFRD diet, serum IL-6 levels were higher in apoE-/-VDR-/- mice than in apoE-/- control animals (Fig 6A). Furthermore, DKO mice exhibited markedly elevated expression of VCAM-1 (Fig 6B) and ICAM-1 (Fig 6C) in the aortae, as well as higher expression of IL-6 than apoE-/- counterparts (Fig 6D). Immunohistochemical staining of the aortic root lesions showed increased expression of VCAM-1 protein, as well as a mild immunoreactivity for ICAM-1 in the vascular endothelium of DKO mice compared to apoE-/- animals (Fig 6G). In the absence of the apoE- genotype and evident lack of macrophage plaque accumulation, VDR-/- mice still exhibited an increased arterial expression of adhesion molecules (Fig 6E and 6F).

Bottom Line: Inhibition of NF-κB activation with super-repressor IκBα blunted all signs of endothelial cell activation caused by downregulation of VDR in endothelial cells.In vivo, deletion of VDR led to significantly larger aortic arch and aortic root lesions in apoE-/- mice, with higher macrophage content. apoE-/-VDR-/-mice showed higher aortic expression of VCAM-1, ICAM-1 and IL-6 when compared to apoE-/-VDR+/+ mice.Our data demonstrate that lack of VDR signaling in endothelial cells leads to a state of endothelial activation with increased leukocyte-endothelial cell interactions that may contribute to the more severe plaque accumulation observed in apoE-/-VDR-/- mice.

View Article: PubMed Central - PubMed

Affiliation: Nephrology Research Department, IRB Lleida, Lleida, Spain.

ABSTRACT
Endothelial cell activation leading to leukocyte recruitment and adhesion plays an essential role in the initiation and progression of atherosclerosis. Vitamin D has cardioprotective actions, while its deficiency is a risk factor for the progression of cardiovascular damage. Our aim was to assess the role of basal levels of vitamin D receptor (VDR) on the early leukocyte recruitment and related endothelial cell-adhesion-molecule expression, as essential prerequisites for the onset of atherosclerosis. Knockdown of VDR in endothelial cells (shVDR) led to endothelial cell activation, characterized by upregulation of VCAM-1, ICAM-1 and IL-6, decreased peripheral blood mononuclear cell (PBMC) rolling velocity and increased PBMC rolling flux and adhesion to the endothelium. shVDR cells showed decreased IκBα levels and accumulation of p65 in the nucleus compared to shRNA controls. Inhibition of NF-κB activation with super-repressor IκBα blunted all signs of endothelial cell activation caused by downregulation of VDR in endothelial cells. In vivo, deletion of VDR led to significantly larger aortic arch and aortic root lesions in apoE-/- mice, with higher macrophage content. apoE-/-VDR-/-mice showed higher aortic expression of VCAM-1, ICAM-1 and IL-6 when compared to apoE-/-VDR+/+ mice. Our data demonstrate that lack of VDR signaling in endothelial cells leads to a state of endothelial activation with increased leukocyte-endothelial cell interactions that may contribute to the more severe plaque accumulation observed in apoE-/-VDR-/- mice. The results reveal an important role for basal levels of endothelial VDR in limiting endothelial cell inflammation and atherosclerosis.

No MeSH data available.


Related in: MedlinePlus