5-aza-2´-Deoxycytidine, a DNA Methyltransferase Inhibitor, Facilitates the Inorganic Phosphorus-Induced Mineralization of Vascular Smooth Muscle Cells

Azechi, Takuya; Sato, Fumiaki; Sudo, Ryo; Wachi, Hiroshi

doi:10.5551/jat.20818

Cited by 36 publications

(29 citation statements)

References 54 publications

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“…Our previous investigation 9) has shown VC involving apoptosis in the ring culture model, which corresponded to the present study, without phenotypic transition of VSMCs. Therefore, this model may represent the passive mineralization of aortic rings induced by high phosphorus or the initial stage of calcification unlike previous studies using cell cult ure model 15,20,21) . …”

Section: Limitationmentioning

confidence: 95%

Mineral Composition of Phosphate-Induced Calcification in a Rat Aortic Tissue Culture Model

Sonou

Ohya

Yashiro

et al. 2015

JAT

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Aim: High phosphorus conditions promote vascular calcification (VC) in both chronic kidney disease (CKD) patients and experimental models. However, the composition of medial calcification has not been accurately determined, so the objective of this study was to evaluate the mineral composition of calcification in a tissue culture model, not a cell culture system. Methods: Aortic rings obtained from male Sprague-Dawley rats were incubated in serum-supplemented medium for 10 days. The inorganic phosphate (Pi) concentration of the medium was increased to induce VC, which was assessed by histology, imaging, and spectroscopy. The mineral composition of the calcification was analyzed using Fourier transform infrared (FTIR) spectroscopic imaging, scanning electron microscopy (SEM), and energy dispersive X-ray spectroscopy (EDX) mapping. Results: The calcium content significantly increased only in aortic rings cultured for 10 days in the high-Pi medium (HiP: 3.8 mmol/L). The concentration of the phosphate transporter Pit-1 in the aortic tissue exposed to HiP was higher than that in the control incubated sections. The FTIR images and spectra indicated that PO4 3 was mostly distributed as hydroxyapatite in the medial calcification of aortic rings cultured in HiP. A small quantity of carbonate was identified. The SEM -EDX overlay map demonstrated that phosphorus and calcium simultaneously accumulated and localized in the area of medial calcification induced by exposure to HiP. Conclusion: This is the first report of accurate determination of the chemical composition of aortic medial calcification. Exposure to high Pi concentration augments aortic calcification via an increase in Pit-1, which mainly contains calcium phosphate.

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Section: Limitationmentioning

confidence: 95%

Mineral Composition of Phosphate-Induced Calcification in a Rat Aortic Tissue Culture Model

Sonou

Ohya

Yashiro

et al. 2015

JAT

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“…Interestingly, high-fat diet or LPS challenge downregulated miR-152, a negative regulator of DNMT1, leading to hypermethylation of the estrogen receptor-α gene in human or rat aortic SMC [57]. Phenotypically modulated SMC can also contribute to vascular calcification, and a recent study reported that inhibition of DNMTs with 5-aza-2′-deoxycytidine (5-Aza) facilitated mineralization in cultured human aortic SMC, likely through demethylation of the alkaline phosphatase promoter [58]. …”

Section: Role Of Epigenetics In Regulating Smc Plasticitymentioning

confidence: 99%

Epigenetic regulation of smooth muscle cell plasticity

Liu

Leslie

Martin

2015

Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanism

120

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Smooth muscle cells (SMC) are the major cell type in blood vessels. Their principle function in the body is to regulate blood flow and pressure through vessel wall contraction and relaxation. Unlike many other mature cell types in the adult body, SMC do not terminally differentiate but retain a remarkable plasticity. They have the unique ability to toggle between a differentiated and quiescent “contractile” state and a highly proliferative and migratory “synthetic” phenotype in response to environmental stresses. While there have been major advances in our understanding of SMC plasticity through the identification of growth factors and signals that can influence the SMC phenotype, how these regulate SMC plasticity remains unknown. To date, several key transcription factors and regulatory cis elements have been identified that play a role in modulating SMC state. The frontier in understanding the molecular mechanisms underlying SMC plasticity has now advanced to the level of epigenetics. This review will summarize the epigenetic regulation of SMC, highlighting the role of histone modification, DNA methylation, and our most recent identification of a DNA demethylation pathway in SMC that is pivotal in the regulation of the SMC phenotypic state. Many disorders are associated with smooth muscle dysfunction, including atherosclerosis, the major underlying cause of stroke and coronary heart disease, as well as transplant vasculopathy, aneurysm, asthma, hypertension, and cancer. An increased understanding of the major regulators of SMC plasticity will lead to the identification of novel target molecules that may, in turn, lead to novel drug discoveries for the treatment of these diseases.

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“…Real-time PCR (qPCR) was carried out using a StepOne Real-Time PCR System (Life Technologies) with a KAPA SYBR â FAST ABI Prism â (Kapa Biosystems, USA), according to the manufacturer's instructions. The primers were described previously Azechi et al 2014) Genes to Cells (2015) 20, 1077-1087…”

Section: Reverse Transcription Polymerase Chain Reactionmentioning

confidence: 99%

MiR‐29‐mediated elastin down‐regulation contributes to inorganic phosphorus‐induced osteoblastic differentiation in vascular smooth muscle cells

et al. 2015

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Vascular calcification increases the risk of cardiovascular mortality. We previously reported that expression of elastin decreases with progression of inorganic phosphorus (Pi)-induced vascular smooth muscle cell (VSMC) calcification. However, the regulatory mechanisms of elastin mRNA expression during vascular calcification remain unclear. MicroRNA-29 family members (miR-29a, b and c) are reported to mediate elastin mRNA expression. Therefore, we aimed to determine the effect of miR-29 on elastin expression and Pi-induced vascular calcification. Calcification of human VSMCs was induced by Pi and evaluated measuring calcium deposition. Pi stimulation promoted Ca deposition and suppressed elastin expression in VSMCs. Knockdown of elastin expression by shRNA also promoted Pi-induced VSMC calcification. Elastin pre-mRNA measurements indicated that Pi stimulation suppressed elastin expression without changing transcriptional activity. Conversely, Pi stimulation increased miR-29a and miR-29b expression. Inhibition of miR-29 recovered elastin expression and suppressed calcification in Pi-treated VSMCs. Furthermore, over-expression of miR-29b promoted Pi-induced VSMC calcification. RT-qPCR analysis showed knockdown of elastin expression in VSMCs induced expression of osteoblast-related genes, similar to Pi stimulation, and recovery of elastin expression by miR-29 inhibition reduced their expression. Our study shows that miR-29-mediated suppression of elastin expression in VSMCs plays a pivotal role in osteoblastic differentiation leading to vascular calcification.

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5-aza-2´-Deoxycytidine, a DNA Methyltransferase Inhibitor, Facilitates the Inorganic Phosphorus-Induced Mineralization of Vascular Smooth Muscle Cells

Cited by 36 publications

References 54 publications

Mineral Composition of Phosphate-Induced Calcification in a Rat Aortic Tissue Culture Model

Mineral Composition of Phosphate-Induced Calcification in a Rat Aortic Tissue Culture Model

Epigenetic regulation of smooth muscle cell plasticity

MiR‐29‐mediated elastin down‐regulation contributes to inorganic phosphorus‐induced osteoblastic differentiation in vascular smooth muscle cells

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