Downregulation of Bone Morphogenetic Protein 4 Expression in Coronary Arterial Endothelial Cells

Gender differences in the development of Pulmonary Artery Hypertension (PAH) have been documented in both human and animal studies. In human, idiopathic PAH is predominantly a disease of young women in their child-bearing years, which suggests a role of female sex hormones in the pathogenesis of PAH. However, the effect of sex hormones on pulmonary vasculatures and the development of PAH has not been fully understood. Recent researches have revealed genetic predisposition such as BMPR (Bone Morphogenetic Protein Receptor). The aim of the present study is to investigate the effect of β-estradiol (E2) and oxygen concentration upon BMPR signal pathway in pulmonary arterial endothelial cells (PAEC) in vitro. Human and rat PAEC were cultured and we examined the expression of BMPR2, BMP-regulated Smads, and Id1 under 21% or 1% O 2 with BMP2 stimulation. Then, we investigate changes in the expression of these molecules in the presence of E2 with or without estrogen receptor antagonist (ICI 182.780.). First, we confirmed that estrogen receptor α and β were expressed in both PAECs. Second, we demonstrated that the expression of mRNA transcripts for BMPR2 and Id1 in PAEC was reduced after exposure to 24 hours’ hypoxia. In addition, E2 decreased the expression of phosphorylated Smad (p-Smad)1/5/8 in a dose-dependent manner (10 −10 M-10 −7 M) and p-Smad1/5/8 expression were decreased about 80% by 10 −7 M of E2. These attenuation of p-Smad1/5/8 expression were rescued by ICI182.780. Third, under normoxic condition with cobalt chloride or deferoxamine to prevent the degradation of HIF (hypoxia-inducible factor)-1α, the presence of E2 decreased the expression of p-Smad1/5/8 like under hypoxia. Conversely, administration of HIF-1α inhibitor (YC-1) canceled the reduced expression of p-Smad1/5/8 like under normoxia. Under hypoxia, the presence of E2 attenuates the BMPR signal pathway in PAEC in vitro. Our data indicated that the advance effect of E2 on BMPR signal pathway was associated with HIF-1α and estrogen receptor. Our observations provide the first evidence that female sex hormone affects on BMPR signal pathway, which can offer new strategies for the treatment of PAH.

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Stimulation of Epithelial Sodium Channels in Endothelial Cells by Bone Morphogenetic Protein-4 Contributes to Salt-Sensitive Hypertension in Rats

Oxidative Medicine and Cellular Longevity ◽

10.1155/2020/3921897 ◽

2020 ◽

Vol 2020 ◽

pp. 1-13

Author(s):

Xu Yang ◽

Na Niu ◽

Chen Liang ◽

Ming-Ming Wu ◽

Liang-Liang Tang ◽

...

Keyword(s):

Endothelial Cells ◽

Bone Morphogenetic Protein ◽

Salt Intake ◽

High Salt ◽

Bone Morphogenetic Protein 4 ◽

Sodium Influx ◽

Morphogenetic Protein ◽

High Salt Intake ◽

Stimulation Of

Previous studies have shown that high salt induces artery stiffness by causing endothelial dysfunction via increased sodium influx. We used our unique split-open artery technique combined with protein biochemistry and in vitro measurement of vascular tone to test a hypothesis that bone morphogenetic protein 4 (BMP4) mediates high salt-induced loss of vascular relaxation by stimulating the epithelial sodium channel (ENaC) in endothelial cells. The data show that high salt intake increased BMP4 both in endothelial cells and in the serum and that exogenous BMP4 stimulated ENaC in endothelial cells. The data also show that the stimulation is mediated by p38 mitogen-activated protein kinases (p38 MAPK) and serum and glucocorticoid-regulated kinase 1 (Sgk1)/neural precursor cell expressed developmentally downregulated gene 4-2 (Nedd4-2) (Sgk1/Nedd4-2). Furthermore, BMP4 decreased mesenteric artery relaxation in a benzamil-sensitive manner. These results suggest that high salt intake stimulates endothelial cells to express and release BMP4 and that the released BMP4 reduces artery relaxation by stimulating ENaC in endothelial cells. Therefore, stimulation of ENaC in endothelial cells by BMP4 may serve as another pathway to participate in the complex mechanism of salt-sensitive (SS) hypertension.

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Bone morphogenetic protein 4 regulates microRNAs miR-494 and miR-126–5p in control of endothelial cell function in angiogenesis

Thrombosis and Haemostasis ◽

10.1160/th16-08-0643 ◽

2017 ◽

Vol 117 (04) ◽

pp. 734-749 ◽

Cited By ~ 17

Author(s):

Erika Saretzki ◽

Franziska Pankratz ◽

Bianca Engert ◽

Sebastian Grundmann ◽

Christoph Bode ◽

...

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Western Blot ◽

Bone Morphogenetic Protein ◽

Cell Function ◽

Umbilical Vein ◽

Endothelial Cell Proliferation ◽

Bone Morphogenetic Protein 4 ◽

Endothelial Cell Function ◽

Morphogenetic Protein

SummaryMicroRNAs are small non-coding RNAs that negatively regulate posttranscriptional gene expression. Several microRNAs have been described to regulate the process of angiogenesis. Previously, we have shown that bone morphogenetic protein 4 (BMP4) increased the proangiogenic activity of endothelial cells. In this project, we now investigated how the pro-angiogenic BMP4 effect is mediated by microRNAs. First, we performed a microRNA array with BMP4-stimulated human umbilical vein endothelial cells (HUVECs). Among the topregulated microRNAs, we detected a decreased expression of miR-494 and increased expression of miR-126–5p. Next, we analysed the canonical Smad and alternative signalling pathways, through which BMP4 would regulate miR-126–5p and miR-494 expression. Furthermore, the functional effect of miR-494 and miR-126–5p on endothelial cells was investigated. MicroRNA-494 overexpression decreased endothelial cell proliferation, migration and sprout formation. Consistently, miR-494 inhibition increased endothelial cell function. As potential miR-494 targets, bFGF and BMP endothelial cell precursorderived regulator (BMPER) were identified and confirmed by western blot. Luciferase assays showed direct miR-494 binding in BMPER 3’UTR. In contrast, miR-126–5p overexpression increased pro-angiogenic endothelial cell behaviour and, accordingly, miR-126–5p inhibition decreased endothelial cell function. As a direct miR-126–5p target we identified the anti-angiogenic thrombospondin-1 which was confirmed by western blot analysis and luciferase assays. In the Matrigel plug assay application of antagomiR-494 increased endothelial cell ingrowth, whereas antagomiR-126–5p treatment decreased cell ingrowth in vivo. Taken together, through differential regulation of the anti-angiomiR-494 and the angiomiR-126–5p by BMP4 both microRNAs contribute to the pro-angiogenic BMP4 effect on endothelial cells.Supplementary Material to this article is available online at www.thrombosis-online.com.

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Drastic Shift From Positive to Negative Estrogen Effect on Bone Morphogenetic Protein Signaling in Pulmonary Arterial Endothelial Cells Under Hypoxia

Circulation Journal ◽

10.1253/circj.cj-12-0997 ◽

2013 ◽

Vol 77 (8) ◽

pp. 2118-2126 ◽

Cited By ~ 6

Author(s):

Hiroaki Ichimori ◽

Shigetoyo Kogaki ◽

Kunihiko Takahashi ◽

Hidekazu Ishida ◽

Jun Narita ◽

...

Keyword(s):

Endothelial Cells ◽

Bone Morphogenetic Protein ◽

Protein Signaling ◽

Bone Morphogenetic Protein Signaling ◽

Morphogenetic Protein ◽

Pulmonary Arterial ◽

Estrogen Effect ◽

Arterial Endothelial Cells

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Bone Morphogenetic Protein 4 Mediates Apoptosis of Capillary Endothelial Cells during Rat Pupillary Membrane Regression

Molecular and Cellular Biology ◽

10.1128/mcb.23.13.4627-4636.2003 ◽

2003 ◽

Vol 23 (13) ◽

pp. 4627-4636 ◽

Cited By ~ 45

Author(s):

Mari Kiyono ◽

Masabumi Shibuya

Keyword(s):

Endothelial Cells ◽

Bone Morphogenetic Protein ◽

Culture System ◽

Neutralizing Antibody ◽

Bone Morphogenetic Protein 4 ◽

Morphogenetic Protein ◽

Molecular Determinants ◽

Capillary Endothelial Cells ◽

Msx Genes ◽

Induced Apoptosis

ABSTRACT Programmed capillary regression is essential for development, but little is known about the mechanism behind this phenomenon. In this study, we characterized the molecular determinants of capillary regression utilizing the pupillary membrane (PM) in the newborn rat's eye. We observed in the 1-day-culture system that apoptotic endothelial cells decrease in number with the addition of a natural antagonist, Noggin, strongly suggesting the involvement of the bone morphogenetic protein (BMP) family in PM regression. In addition, the lens-conditioned medium (Lens-CM) induced apoptosis of HUVE cells and inhibited endothelial tubulogenesis, which were completely blocked by both Noggin and the BMP4-specific neutralizing antibody. Activation of BMP4 pathway in endothelial cells was confirmed by both the up-regulation of Msx genes correlated with apoptosis and the translocation of Smad1 into the nucleus. We showed a transient expression of BMP4 in Lens-CM by immunoprecipitation assay. Furthermore, the transcorneal injection of BMP4 in rats enhanced the apoptosis of PMs, while that of Noggin attenuated it. These results indicate that BMP4 pathways play pivotal roles in capillary regression in a paracrine manner between lens and PMs.

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Ectopic transient overexpression of OCT-4 facilitates BMP4-induced osteogenic transdifferentiation of human umbilical vein endothelial cells

Journal of Tissue Engineering ◽

10.1177/2041731420909208 ◽

2020 ◽

Vol 11 ◽

pp. 204173142090920

Author(s):

Seung Hyun L Kim ◽

Seunghun S Lee ◽

Inseon Kim ◽

Janet Kwon ◽

Song Kwon ◽

...

Keyword(s):

Tissue Engineering ◽

Endothelial Cells ◽

Cell Therapy ◽

Bone Morphogenetic Protein ◽

Umbilical Vein ◽

Cell Types ◽

Bone Morphogenetic Protein 4 ◽

Morphogenetic Protein ◽

Umbilical Vein Endothelial Cells

Limitation in cell sources for autologous cell therapy has been a recent focus in stem cell therapy and tissue engineering. Among various research advances, direct conversion, or transdifferentiation, is a notable and feasible strategy for the generation and acquirement of wanted cell source. So far, utilizing cell transdifferentiation technology in tissue engineering was mainly restricted at achieving single wanted cell type from diverse cell types with high efficiency. However, regeneration of a complete tissue always requires multiple cell types which poses an intrinsic complexity. In this study, enhanced osteogenic differentiation was achieved by transient ectopic expression of octamer-binding transcription factor 4 ( OCT-4) gene followed by bone morphogenetic protein 4 treatment on human umbilical vein endothelial cells. OCT-4 transfection and bone morphogenetic protein 4 treatment resulted in enhanced expression of osteogenic markers such as core-binding factor alpha 1, alkaline phosphatase, and collagen 1 compared with bone morphogenetic protein 4 treatment alone. Furthermore, we employed gelatin-heparin cryogel in cranial defect model for in vivo bone formation. Micro-computed tomography and histological analysis of in vivo samples showed that OCT-4 transfection followed by bone morphogenetic protein 4 treatment resulted in efficient transdifferentiation of endothelial cells to osteogenic cells. These results suggest that the combination of OCT-4 and bone morphogenetic protein 4 on endothelial cells would be a reliable multicellular transdifferentiation model which could be applied for bone tissue engineering.

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Regulation of Bone Morphogenetic Protein-4 by Matrix GLA Protein in Vascular Endothelial Cells Involves Activin-like Kinase Receptor 1

Journal of Biological Chemistry ◽

10.1074/jbc.m604239200 ◽

2006 ◽

Vol 281 (45) ◽

pp. 33921-33930 ◽

Cited By ~ 83

Author(s):

Yucheng Yao ◽

Amina F. Zebboudj ◽

Esther Shao ◽

Martin Perez ◽

Kristina Boström

Keyword(s):

Endothelial Cells ◽

Bone Morphogenetic Protein ◽

Vascular Endothelial Cells ◽

Matrix Gla Protein ◽

Bone Morphogenetic Protein 4 ◽

Vascular Endothelial ◽

Morphogenetic Protein

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Differential proinflammatory and prooxidant effects of bone morphogenetic protein-4 in coronary and pulmonary arterial endothelial cells

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00180.2008 ◽

2008 ◽

Vol 295 (2) ◽

pp. H569-H577 ◽

Cited By ~ 52

Author(s):

Anna Csiszar ◽

Nazar Labinskyy ◽

Hanjoong Jo ◽

Praveen Ballabh ◽

Zoltan Ungvari

Keyword(s):

Endothelial Cells ◽

Adverse Effects ◽

Bone Morphogenetic Protein ◽

Pulmonary Arteries ◽

Endothelial Activation ◽

Systemic Circulation ◽

Morphogenetic Protein ◽

Pathophysiological Role ◽

Pulmonary Arterial ◽

Arterial Endothelial Cells

There is increasing evidence that TGF-β family member cytokine bone morphogenetic protein (BMP)-4 plays different pathophysiological roles in the pulmonary and systemic circulation. Upregulation of BMP-4 has been linked to atherosclerosis and hypertension in the systemic circulation, whereas disruption of BMP-4 signaling is associated with the development of pulmonary hypertension. To test the hypothesis that BMP-4 elicits differential effects in the pulmonary and systemic circulation, we compared the prooxidant and proinflammatory effects of BMP-4 in cultured human coronary arterial endothelial cells (CAECs) and pulmonary arterial endothelial cells (PAECs). We found that BMP-4 (from 0.3 to 10 ng/ml) in CAECs increased O2•− and H2O2 generation, induced NF-κB activation, upregulated ICAM-1, and induced monocyte adhesiveness to ECs. In contrast, BMP-4 failed to induce oxidative stress or endothelial activation in PAECs. Also, BMP-4 treatment impaired acetylcholine-induced relaxation and increased O2•− production in cultured rat carotid arteries, whereas cultured rat pulmonary arteries were protected from these adverse effects of BMP-4. Thus, we propose that BMP-4 exerts prooxidant, prohypertensive, and proinflammatory effects only in the systemic circulation, whereas pulmonary arteries are protected from these adverse effects of BMP-4. The vascular bed-specific endothelial effects of BMP-4 are likely to contribute to its differential pathophysiological role in the systemic and pulmonary circulation.

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