scholarly journals Adenosine upregulates VEGF expression in cultured myocardial vascular smooth muscle cells

1999 ◽  
Vol 277 (2) ◽  
pp. H595-H602 ◽  
Author(s):  
Jian-Wei Gu ◽  
Ann L. Brady ◽  
Vivek Anand ◽  
Michael C. Moore ◽  
Whitney C. Kelly ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Mrna Expression ◽  
Vascular Smooth Muscle Cells ◽  
Vegf Expression ◽  
Vegf Mrna ◽  
Protein Levels ◽  
Hypoxic Conditions ◽  
Vegf Mrna Expression ◽  
Vegf Protein

We tested whether adenosine has differential effects on vascular endothelial growth factor (VEGF) expression under normoxic and hypoxic conditions, and whether A1 or A2 receptors (A1R; A2R) mediate these effects. Myocardial vascular smooth muscle cells (MVSMCs) from dog coronary artery were exposed to hypoxia (1% O2) or normoxia (20% O2) in the absence and presence of adenosine agonists or antagonists for 18 h. VEGF protein levels were measured in media with ELISA. VEGF mRNA expression was determined with Northern blot analysis. Under normoxic conditions, the adenosine A1R agonists, N 6-cyclopentyladenosine and R(-)- N 6-(2-phenylisopropyl)adenosine did not increase VEGF protein levels at A1R stimulatory concentrations. However, adenosine (5 μM) and the adenosine A2R agonist N 6-[2-(3,5-dimethoxyphenyl)-2-(2-methylphenyl)]ethyl adenosine (DPMA; 100 nM) increased VEGF protein levels by 51 and 132% and increased VEGF mRNA expression by 44 and 90%, respectively, in cultured MVSMCs under normoxic conditions. Hypoxia caused an approximately fourfold increase in VEGF protein and mRNA expression, which could not be augmented with exogenous adenosine, A2R agonist (DPMA), or A1R agonist [1,3-diethyl-8-phenylxanthine (DPX)]. The A2R antagonist 8-(3-chlorostyryl)-caffeine completely blocked adenosine-induced VEGF protein and mRNA expression and decreased baseline VEGF protein levels by up to ∼60% under normoxic conditions but only by ∼25% under hypoxic conditions. The A1R antagonist DPX had no effect. These results are consistent with the hypothesis that 1) adenosine increases VEGF protein and mRNA expression by way of A2R. 2) Adenosine plays a major role as an autocrine factor regulating VEGF expression during normoxic conditions but has a relatively minor role during hypoxic conditions. 3) Endogenous adenosine can account for the majority of basal VEGF secretion by MVSMCs under normoxic conditions and could therefore be a maintenance factor for the vasculature.

Laser-assisted microdissection and real-time PCR detect anti-inflammatory effect of perfluorocarbon

2003 ◽  
Vol 285 (1) ◽  
pp. L55-L62 ◽  
Author(s):  
Katharina von der Hardt ◽  
Michael Andreas Kandler ◽  
Ludger Fink ◽  
Ellen Schoof ◽  
Jörg Dötsch ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Mrna Expression ◽  
Vascular Smooth Muscle Cells ◽  
Alveolar Macrophages ◽  
Real Time Pcr ◽  
Muscle Cells ◽  
Cell Types ◽  
Anti Inflammatory ◽  
Inflammatory Effect

The aim of this study was to identify cell types involved in the anti-inflammatory effect of ventilation with perfluorocarbon in vivo. Fifteen anesthetized, surfactant-depleted piglets received either aerosolized perfluorocarbon (Aerosol-PFC), partial liquid ventilation (rLV) at functional residual capacity (FRC) volume (FRC-PLV), or intermittent mandatory ventilation (control). After laser-assisted microdissection of different lung cell types, mRNA expression of IL-8 and ICAM-1 was determined using TaqMan real-time PCR normalized to hypoxanthine phosphoribosyltransferase (HPRT). IL-8 mRNA expression (means ± SE; control vs. Aerosol-PFC) was 356 ± 142 copies IL-8 mRNA/copy HPRT mRNA vs. 3.5 ± 1.8 in alveolar macrophages ( P <0.01); 208 ± 108 vs. 2.7 ± 0.8 in bronchiolar epithelial cells ( P <0.05); 26 ± 11 vs. 0.7 ± 0.2 in alveolar septum cells ( P <0.01); 2.8 ± 1.0 vs. 0.8 ± 0.4 in bronchiolar smooth muscle cells ( P <0.05); and 1.1 ± 0.4 vs. 0.2 ± 0.05 in vascular smooth muscle cells ( P <0.05). With FRC-PLV, IL-8/HPRT mRNA expression was significantly lower in macrophages, bronchiolar epithelial, and vascular smooth muscle cells. ICAM-1 mRNA expression in vascular endothelial cells remained unchanged. Predominantly, alveolar macrophages and bronchiolar epithelial cells were involved in the inflammatory pulmonary process. The anti-inflammatory effect of Aerosol-PFC was most pronounced.

Hypoxia-induced expression of VEGF is reversible in myocardial vascular smooth muscle cells

1997 ◽  
Vol 273 (2) ◽  
pp. H628-H633 ◽  
Author(s):  
J. W. Gu ◽  
T. H. Adair
Keyword(s):  
Smooth Muscle ◽  
Growth Factor ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Conditioned Media ◽  
Dependent Manner ◽  
Induced Expression ◽  
Protein Levels

We determined whether hypoxia-induced expression of vascular endothelial growth factor (VEGF) can be reversed by a normoxic environment. Dog myocardial vascular smooth muscle cells (MVSMCs) were exposed to hypoxia (1% O2) for 24 h and then returned to normoxia (20% O2). VEGF protein levels increased by more than fivefold after 24 h of hypoxia and returned to baseline within 24 h of the return of the cells to normoxia. Northern blot analysis showed that hypoxia caused a 5.5-fold increase in VEGF mRNA, and, again, the expression was reversed after reinstitution of normoxia. Additional measurements showed that basic fibroblast growth factor and platelet-derived growth factor protein levels were not induced by hypoxia and that hypoxia caused a fourfold decrease in transforming growth factor-beta 1 protein levels. Hypoxia conditioned media from MVSMCs caused human umbilical vein endothelial cells to increase [3H]thymidine incorporation by twofold, an effect that was blocked in a dose-dependent manner by anti-human VEGF antibody. The hypoxia conditioned media had no effect on MVSMC proliferation. These findings suggest that VEGF expression can be bidirectionally controlled by tissue oxygenation, and thus support the hypothesis that VEGF is a physiological regulator of angiogenesis.

Cyclooxygenase-2 induction by bradykinin in aortic vascular smooth muscle cells

2006 ◽  
Vol 290 (1) ◽  
pp. H30-H36 ◽  
Author(s):  
Jorge A. Rodriguez ◽  
Paula De la Cerda ◽  
Eileen Collyer ◽  
Valerie Decap ◽  
Carlos P. Vio ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Receptor Antagonist ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Receptor Activation ◽  
Atheromatous Plaque ◽  
Protein Levels ◽  
Cox 2

Vascular smooth muscle cell proliferation and migration play an important role in the pathophysiology of several vascular diseases, including atherosclerosis. Prostaglandins that have been implicated in this process are synthesized by two isoforms of cyclooxygenase (COX), with the expression of the regulated COX-2 isoform increased in atherosclerotic plaques. Bradykinin (BK), a vasoactive peptide increased in inflammation, induces the formation of prostaglandins through specific receptor activation. We hypothesized that BK plays an important role in the regulation of COX-2, contributing to the increase in production of prostaglandins in vascular smooth muscle cells. Herein we examined the signaling pathways that participate in the BK regulation of COX-2 protein levels in primary cultured aortic vascular smooth muscle cells. We observed an increase in COX-2 protein levels induced by BK that was maximal at 24 h. This increase was blocked by a B2 kinin receptor antagonist but not a B1 receptor antagonist, suggesting that the B2 receptor is involved in this pathway. In addition, we conclude that the activation of mitogen-activated protein kinases p42/p44, protein kinase C, and nitric oxide synthase is necessary for the increase in COX-2 levels induced by BK because either of the specific inhibitors for these enzymes blocked the effect of BK. Using a similar approach, we further demonstrated that reactive oxygen species and cAMP were not mediators on this pathway. These results suggest that BK activates several intracellular pathways that act in combination to increase COX-2 protein levels. This study suggests a role for BK on the evolution of the atheromatous plaque by virtue of controlling the levels of COX-2.

Inducibleendothelin mRNA expression and peptide secretion in cultured human vascular smooth muscle cells

1990 ◽  
Vol 168 (3) ◽  
pp. 1303-1310 ◽  
Author(s):  
Thérèse J. Resink ◽  
Alfred W.A. Hahn ◽  
Timothy Scott-Burden ◽  
Jerry Powell ◽  
Erika Weber ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Mrna Expression ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
Peptide Secretion

scholarly journals Heart and lung VEGF mRNA expression in rats with monocrotaline- or hypoxia-induced pulmonary hypertension

1998 ◽  
Vol 275 (6) ◽  
pp. H1948-H1956 ◽  
Author(s):  
Chohreh Partovian ◽  
Serge Adnot ◽  
Saadia Eddahibi ◽  
Emmanuel Teiger ◽  
Micheline Levame ◽  
...  
Keyword(s):  
Pulmonary Hypertension ◽  
Mrna Expression ◽  
Vascular Remodeling ◽  
Prolonged Exposure ◽  
Similar Degree ◽  
Vegf Expression ◽  
Vegf Mrna ◽  
Pulmonary Vascular Remodeling ◽  
The Right ◽  
Vegf Mrna Expression

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific mitogen that is upregulated during exposure to hypoxia. In this study, we analyzed heart and lung VEGF mRNA expression and examined pulmonary vascular remodeling as well as myocardial capillary density in two rat models of pulmonary hypertension involving exposure to chronic hypoxia (CH) and treatment with monocrotaline (MCT), respectively. The rats were studied after 0.5, 1, 3, 15, and 30 days of exposure to 10% O2 or 1, 6, and 30 days after a subcutaneous MCT injection (60 mg/kg). Both CH and MCT induced pulmonary hypertension and hypertrophy of the right ventricle (RV) with increased RV weight and atrial natriuretic peptide mRNA expression. VEGF mRNA expression as assessed by Northern blot analysis was potently induced after 12 h of hypoxia in both the right and left ventricles. After prolonged exposure to hypoxia, VEGF mRNA returned to baseline in the left ventricle (LV) but remained increased in the RV, where it peaked after 30 days. In MCT rats, VEGF mRNA was unchanged in the LV but decreased by 50% in the RV and by 90% in the lungs after 30 days. VEGF mRNA remained unchanged in the lungs from CH rats. Pulmonary vascular remodeling was more pronounced in MCT than in CH rats. The number of capillaries per RV myocyte was increased in rats exposed to 30 days of hypoxia, whereas it remained unchanged in MCT rats despite a similar degree of RV hypertrophy. Our results suggest that the sustained increase in VEGF expression in the hypertrophied RV during CH may account for the increased number of capillaries per myocyte. In contrast, reduced VEGF expression in the lungs and RV of MCT rats may aggravate pulmonary vascular remodeling and compromise RV myocardial perfusion.

Abstract 544: Diabetes Mellitus Impacts on Vascular Calcification in Peripheral Arterial Disease Patients but Does Not Further Activates Osteogenic Markers in Beta-Glycerophosphate Stimulated Vascular Smooth Muscle Cells

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Elisangela Farias-Silva ◽  
Cynthia A Mendes ◽  
Luciana A Pescatore ◽  
Maria Claudina C Andrade ◽  
Luciana S Carmo ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Smooth Muscle ◽  
Peripheral Arterial Disease ◽  
Protein Expression ◽  
Smooth Muscle Cells ◽  
Mrna Expression ◽  
Vascular Calcification ◽  
Vascular Smooth Muscle Cells ◽  
Arterial Disease ◽  
Peripheral Arterial

Introduction: Diabetes mellitus (DM) accelerates vascular calcification (VC) in peripheral arterial disease (PAD), increasing limb ischemia and amputation risk. Although DM and VC implicates in PAD, the mechanisms underlying vascular smooth muscle cells (SMC) osteochondrogenic differentiation in this setting are scarce. Objectives: To assess VC and osteogenic mRNA/protein expression (Msx2, Runx2 and ALPL) in arteries and in primary SMC isolated from PAD patients with DM (n=3), without DM (n=2) and individuals without PAD (control [CTL], n=2) that underwent amputation. Methods: Ethical committee approved study and patients signed informed consent. 5-7 th passages SMC were incubated without (untreated) or with β-glycerophosphate 10mM (β-GP) for 48h (mRNA expression), 72h (protein expression) and 14 days (calcification). In addition, we performed tissue immunofluorescence and Alizarin Red S. Results: Untreated PAD+DM SMC increased calcification (4.6±1.2) in comparison to PAD without DM patients (2.5±0.2) and CTL individuals (1.0±0.07). β-GP further increased calcification in respective groups (51.3±4.7; 13.5±1.1 and 9.8±1.2). Msx2 mRNA expression decreased in CTL SMC after β-GP (48h) and did not change in PAD without DM and in PAD+DM SMC. Msx2 protein expression decreased in all groups after calcifying medium (72h). Moreover, Runx2 mRNA expression increased in β-GP-treated PAD+DM SMC and ALPL mRNA expression augmented in β-GP-treated SMC from PAD without DM patients. However, Runx2 and ALPL protein expression was not modulated by calcifying medium (72h) in all groups. Interestingly, immunofluorescence of arterial samples demonstrated increased osteochondrogenic protein expression (Msx2, Runx2 and ALPL) around calcifying foci in PAD+DM group (+++) versus PAD without DM (++) and versus CTL (+) patients. Coincidently, PAD+DM group showed augmented vascular calcification (19x10 4 ±10x10 4 ; 7x10 4 ±2x10 4 ; 3x10 3 ±10 -2 μm 2 ). Conclusion: We demonstrated increased calcification in arteries from amputated PAD+DM patients and in respective SMC without and with calcifying medium. Although we found augmented osteochondrogenic protein expression in the vessel wall from PAD+DM individuals this was not robustly shown in isolated SMC in vitro .

Abstract 439: Biomechanical Stimulation Promotes Translocation of Nuclear Factor of Activated T-cells 5 Into The Nucleus of Vascular Smooth Muscle Cells

2015 ◽  
Vol 35 (suppl_1) ◽  
Author(s):  
Maren Hödebeck ◽  
Markus Hecker ◽  
Thomas Korff
Keyword(s):  
T Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Mrna Expression ◽  
Vascular Smooth Muscle Cells ◽  
Splice Variant ◽  
Nuclear Translocation ◽  
Nuclear Factor ◽  
Activated T Cells ◽  
Biomechanical Stimulation

Chronic alterations in the biomechanical stimulation of vascular smooth muscle cells (VSMC) as experienced during hypertension lead to changes in VSMC phenotype and function and further enable the structural remodeling of the vessel wall. In this context, we recently reported that an increase in wall stress or biomechanical stretch is sufficient to activate nuclear factor of activated T-cells 5 (NFAT5). This transcription factor promotes the expression of gene products such as tenascin-C and κ-actin, both involved in VSMC migration. Based on these findings, we hypothesized that biomechanical stretch elicits NFAT5 mRNA expression and induces biochemical modifications of NFAT5 on the post-translational level, a prerequisite for its entry into the nucleus and transcriptional activity. To scrutinize this hypothesis, human arterial VSMC were exposed to biomechanical stretch (13%, 0.5 Hz) and subjected to detailed mRNA expression analyses. While a ~3-fold reduction in NFAT5 splice variant 1 (isoform A) mRNA expression was observed in stretch-stimulated VSMC as compared to the static controls (n=3, p<0.05), splice variant 3 (isoform C) mRNA levels were induced ~1.8-fold (n=3, p<0.05). Overexpression of corresponding Flag-tagged NFAT5 proteins in VSMC and subsequent immunofluorescence as well as biochemical analyses revealed that isoform A was primarily located in the cytoplasm of static and stretch-stimulated VSMC while isoform C was preferentially localized in the nucleus under baseline conditions and further accumulated in the nucleus upon biomechanical stimulation (n=3, p<0.05). Nuclear translocation of isoform C was amplified for phosphorylation-deficient mutants generated by exchanging serine to alanine at position 1197 even under static culture conditions while a phosphomimetic mutation at this residue (serine to glutamate) inhibited NFAT5c nuclear translocation (n=3, p<0.05). Collectively, our findings indicate that exposure of VSMC to biomechanical stretch triggers the expression of NFAT5 isoform C and controls its entry into the nucleus via phosphorylation at S-1197. Current investigations are focusing on the impact of NFAT5 on hypertensive remodeling utilizing inducible smooth muscle cell-specific NFAT5-deficient mice.

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