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1,25 Dihydroxyvitamin D3 Inhibits TGFβ1-Mediated Primary Human Cardiac Myofibroblast Activation.

Meredith A, Boroomand S, Carthy J, Luo Z, McManus B - PLoS ONE (2015)

Bottom Line: Functional responses of fibroblasts were analyzed by in vitro collagen gel contraction assay. 1,25(OH)2D3 treatment significantly inhibited TGFβ1-mediated cell contraction, and confocal imaging demonstrated reduced stress fiber formation in the presence of 1,25(OH)2D3.Treatment with 1,25(OH)2D3 reduced alpha-smooth muscle actin expression to control levels and inhibited SMAD2 phosphorylation.Our results demonstrate that active vitamin D can prevent TGFβ1-mediated biochemical and functional pro-fibrotic changes in human primary cardiac fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Centre for Heart Lung Innovation, St. Paul's Hospital, University of British Columbia, Vancouver, BC, Canada; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.

ABSTRACT

Aims: Epidemiological and interventional studies have suggested a protective role for vitamin D in cardiovascular disease, and basic research has implicated vitamin D as a potential inhibitor of fibrosis in a number of organ systems; yet little is known regarding direct effects of vitamin D on human cardiac cells. Given the critical role of fibrotic responses in end stage cardiac disease, we examined the effect of active vitamin D treatment on fibrotic responses in primary human adult ventricular cardiac fibroblasts (HCF-av), and investigated the relationship between circulating vitamin D (25(OH)D3) and cardiac fibrosis in human myocardial samples.

Methods and results: Interstitial cardiac fibrosis in end stage HF was evaluated by image analysis of picrosirius red stained myocardial sections. Serum 25(OH)D3 levels were assayed using mass spectrometry. Commercially available HCF-av were treated with transforming growth factor (TGF)β1 to induce activation, in the presence or absence of active vitamin D (1,25(OH)2D3). Functional responses of fibroblasts were analyzed by in vitro collagen gel contraction assay. 1,25(OH)2D3 treatment significantly inhibited TGFβ1-mediated cell contraction, and confocal imaging demonstrated reduced stress fiber formation in the presence of 1,25(OH)2D3. Treatment with 1,25(OH)2D3 reduced alpha-smooth muscle actin expression to control levels and inhibited SMAD2 phosphorylation.

Conclusions: Our results demonstrate that active vitamin D can prevent TGFβ1-mediated biochemical and functional pro-fibrotic changes in human primary cardiac fibroblasts. An inverse relationship between vitamin D status and cardiac fibrosis in end stage heart failure was observed. Collectively, our data support an inhibitory role for vitamin D in cardiac fibrosis.

No MeSH data available.


Related in: MedlinePlus

Vitamin D does not inhibit TGFβ1-mediated cellular proliferation.Evaluation of proliferation rates in our treatment groups revealed no significant change in cellular proliferation between cells treated with active vitamin D and TGFβ1 or TGFβ1 alone. Proliferation was increased in the presence of TGFβ1. All data represent mean ± SEM. p-values were calculated using one way analysis of variance with a Bonferroni multiple comparison test.
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pone.0128655.g003: Vitamin D does not inhibit TGFβ1-mediated cellular proliferation.Evaluation of proliferation rates in our treatment groups revealed no significant change in cellular proliferation between cells treated with active vitamin D and TGFβ1 or TGFβ1 alone. Proliferation was increased in the presence of TGFβ1. All data represent mean ± SEM. p-values were calculated using one way analysis of variance with a Bonferroni multiple comparison test.

Mentions: The biochemical changes elicited by TGFβ1 are paralleled by functional changes in vitro, mediated by the contractile phenotype of activated myofibroblasts. To examine whether vitamin D can act to inhibit these functional effects we performed a collagen gel contraction assay. Fibroblast contraction, as measured by this model, was significantly increased in cells treated with TGFβ1 at all timepoints. Concurrent treatment with TGFβ1 + 1,25(OH)2D3 reduced collagen gel contraction to untreated control levels throughout the 96 hour time course (p<0.05) (Fig 2E). To determine whether the changes in gel contraction we observed resulted from increased cellular proliferation in the presence of TGFβ1 we performed a BrdU incorporation proliferation assay. Although TGFβ1 significantly increased cell numbers, there was no significant difference in cells co-treated with 1,25(OH)2D3 (Fig 3) suggesting the observed differences in gel contraction were not a function of cell number.


1,25 Dihydroxyvitamin D3 Inhibits TGFβ1-Mediated Primary Human Cardiac Myofibroblast Activation.

Meredith A, Boroomand S, Carthy J, Luo Z, McManus B - PLoS ONE (2015)

Vitamin D does not inhibit TGFβ1-mediated cellular proliferation.Evaluation of proliferation rates in our treatment groups revealed no significant change in cellular proliferation between cells treated with active vitamin D and TGFβ1 or TGFβ1 alone. Proliferation was increased in the presence of TGFβ1. All data represent mean ± SEM. p-values were calculated using one way analysis of variance with a Bonferroni multiple comparison test.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4462580&req=5

pone.0128655.g003: Vitamin D does not inhibit TGFβ1-mediated cellular proliferation.Evaluation of proliferation rates in our treatment groups revealed no significant change in cellular proliferation between cells treated with active vitamin D and TGFβ1 or TGFβ1 alone. Proliferation was increased in the presence of TGFβ1. All data represent mean ± SEM. p-values were calculated using one way analysis of variance with a Bonferroni multiple comparison test.
Mentions: The biochemical changes elicited by TGFβ1 are paralleled by functional changes in vitro, mediated by the contractile phenotype of activated myofibroblasts. To examine whether vitamin D can act to inhibit these functional effects we performed a collagen gel contraction assay. Fibroblast contraction, as measured by this model, was significantly increased in cells treated with TGFβ1 at all timepoints. Concurrent treatment with TGFβ1 + 1,25(OH)2D3 reduced collagen gel contraction to untreated control levels throughout the 96 hour time course (p<0.05) (Fig 2E). To determine whether the changes in gel contraction we observed resulted from increased cellular proliferation in the presence of TGFβ1 we performed a BrdU incorporation proliferation assay. Although TGFβ1 significantly increased cell numbers, there was no significant difference in cells co-treated with 1,25(OH)2D3 (Fig 3) suggesting the observed differences in gel contraction were not a function of cell number.

Bottom Line: Functional responses of fibroblasts were analyzed by in vitro collagen gel contraction assay. 1,25(OH)2D3 treatment significantly inhibited TGFβ1-mediated cell contraction, and confocal imaging demonstrated reduced stress fiber formation in the presence of 1,25(OH)2D3.Treatment with 1,25(OH)2D3 reduced alpha-smooth muscle actin expression to control levels and inhibited SMAD2 phosphorylation.Our results demonstrate that active vitamin D can prevent TGFβ1-mediated biochemical and functional pro-fibrotic changes in human primary cardiac fibroblasts.

View Article: PubMed Central - PubMed

Affiliation: Centre for Heart Lung Innovation, St. Paul's Hospital, University of British Columbia, Vancouver, BC, Canada; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.

ABSTRACT

Aims: Epidemiological and interventional studies have suggested a protective role for vitamin D in cardiovascular disease, and basic research has implicated vitamin D as a potential inhibitor of fibrosis in a number of organ systems; yet little is known regarding direct effects of vitamin D on human cardiac cells. Given the critical role of fibrotic responses in end stage cardiac disease, we examined the effect of active vitamin D treatment on fibrotic responses in primary human adult ventricular cardiac fibroblasts (HCF-av), and investigated the relationship between circulating vitamin D (25(OH)D3) and cardiac fibrosis in human myocardial samples.

Methods and results: Interstitial cardiac fibrosis in end stage HF was evaluated by image analysis of picrosirius red stained myocardial sections. Serum 25(OH)D3 levels were assayed using mass spectrometry. Commercially available HCF-av were treated with transforming growth factor (TGF)β1 to induce activation, in the presence or absence of active vitamin D (1,25(OH)2D3). Functional responses of fibroblasts were analyzed by in vitro collagen gel contraction assay. 1,25(OH)2D3 treatment significantly inhibited TGFβ1-mediated cell contraction, and confocal imaging demonstrated reduced stress fiber formation in the presence of 1,25(OH)2D3. Treatment with 1,25(OH)2D3 reduced alpha-smooth muscle actin expression to control levels and inhibited SMAD2 phosphorylation.

Conclusions: Our results demonstrate that active vitamin D can prevent TGFβ1-mediated biochemical and functional pro-fibrotic changes in human primary cardiac fibroblasts. An inverse relationship between vitamin D status and cardiac fibrosis in end stage heart failure was observed. Collectively, our data support an inhibitory role for vitamin D in cardiac fibrosis.

No MeSH data available.


Related in: MedlinePlus