Nitric oxide stimulates myoglobin gene and protein expression in vascular smooth muscle

2009 ◽  
Vol 423 (2) ◽  
pp. 169-177 ◽  
Author(s):  
Benjamin S. Rayner ◽  
Susan Hua ◽  
Tharani Sabaretnam ◽  
Paul K. Witting
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Cell Viability ◽  
Transcriptional Activation ◽  
Cell Signalling ◽  
No Production ◽  
Parallel Increase ◽  
Signalling Molecule ◽  
Gene And Protein Expression

Mb (myoglobin) is a haemoprotein present in cardiac, skeletal and smooth muscle and is primarily responsible for the storage and ‘facilitated transfer’ of molecular oxygen from the cell membrane to mitochondria. Also, Mb plays a role in regulating •NO (nitric oxide) homoeostasis through (i) binding •NO (Mb–NO complex); (ii) oxidation of •NO to nitrate; and (iii) formation of vasoactive S-nitroso-Mb [Rayner, B.S., Wu, B.-J., Raftery, M., Stocker, R. and Witting, P.K. (2005) J. Biol. Chem. 280, 9985–9993]. Pathological •NO concentrations affect mitochondrial function and decrease cell viability through inducing apoptosis. Treatment of cultured rat VSMCs (vascular smooth muscle cells) with cumulative doses (0.1, 1 or 10 μM) of •NO from the donors diethylamineNONOate or spermineNONOate (N-[2-aminoethyl]-N-[2-hydroxy-3-nitrosohydrazine]-1,2-ethelenediamine) yielded a time-dependent increase in Mb gene expression. Concomitant transcriptional activation increased the concentration of Mb within cultured rat or primary human VSMCs as judged by Western blot analysis and indirect immunofluorescence microscopy. Cell viability did not decrease in these cells at the •NO doses tested. Importantly, sub-culturing isolated rat aortic segments for 7 days in the presence of L-arginine at 37 °C stimulated •NO production with a parallel increase in Mb in the underlying VSMCs. Overall, exposure of VSMCs (either in cell culture or intact vessels) to pathological •NO promotes an up-regulation of the Mb gene and protein, suggesting a feedback relationship between •NO and Mb that regulates the concentration of the potent cell signalling molecule in the vessel wall, similar to the role haemoglobin plays in the vessel lumen.

TNF-alpha and IFN-gamma induce expression of nitric oxide synthase in cultured rat medullary interstitial cells

1995 ◽  
Vol 269 (2) ◽  
pp. F212-F217 ◽  
Author(s):  
K. S. Lau ◽  
O. Nakashima ◽  
G. R. Aalund ◽  
L. Hogarth ◽  
K. Ujiie ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Nitric Oxide Synthase ◽  
Vascular Smooth Muscle ◽  
Guanylyl Cyclase ◽  
Soluble Guanylyl Cyclase ◽  
Tnf Alpha ◽  
No Production ◽  
Ifn Gamma ◽  
Oxide Synthase

Cytokines increase the expression of the inducible (type II) nitric oxide synthase (NOS) in macrophages, liver, and renal epithelial cells. Previously, we found that cultured rat medullary interstitial cells (RMIC) contain high levels of soluble guanylyl cyclase. To determine whether these cells can also produce NO, we studied the effects of tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) on NO production, NOS II mRNA, and NOS II protein expression. Both TNF-alpha and IFN-gamma, in the presence of a low concentration of the other cytokine, caused dose-dependent increases in NO production. Exposure to TNF-alpha and IFN-gamma stimulated the production of NOS II mRNA, as determined by Northern blotting. Restriction mapping of reverse transcription-polymerase chain reaction products indicated that normal cells contained macrophage NOS II, whereas cytokine-stimulated cells contained primarily vascular smooth muscle NOS II and some macrophage NOS II. The appearance of NOS II protein was demonstrated by Western blotting. RMIC cell guanosine 3',5'-cyclic monophosphate accumulation increased 129-fold in response to the cytokines. NOS inhibitors decreased nitrite production. We conclude that 1) TNF-alpha and IFN-gamma induce the expression of vascular smooth muscle NOS II and production of NO in RMIC, and 2) NO acts as an autocrine activator of the soluble guanylyl cyclase in RMIC.

scholarly journals Complex interactions of NO/cGMP/PKG systems on Ca2+ signaling in afferent arteriolar vascular smooth muscle

2010 ◽  
Vol 298 (1) ◽  
pp. H144-H151 ◽  
Author(s):  
Susan K. Fellner ◽  
William J. Arendshorst
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
No Production ◽  
No Donor ◽  
Complex Interactions ◽  
Resistance Vessels ◽  
Microsphere Method ◽  
Afferent Arterioles ◽  
Oxide Synthase

Little is known about the effects of nitric oxide (NO) and the cyclic GMP (cGMP)/protein kinase G (PKG) system on Ca2+ signaling in vascular smooth muscle cells (VSMC) of resistance vessels in general and afferent arterioles in particular. We tested the hypotheses that cGMP-, Ca2+-dependent big potassium channels (BKCa2+) buffer the Ca2+ response to depolarization by high extracellular KCl and that NO inhibits adenosine diphosphoribose (ADPR) cyclase, thereby reducing the Ca2+-induced Ca2+ release. We isolated rat afferent arterioles, utilizing the magnetized microsphere method, and measured cytosolic Ca2+ concentration ([Ca2+]i) with fura-2, a preparation in which endothelial cells do not participate in [Ca2+]i responses. KCl (50 mM)-induced depolarization causes an immediate increase in [Ca2+]i of 151 nM. The blockers Nω-nitro-l-arginine methyl ester (of nitric oxide synthase), 1,2,4-oxodiazolo-[4,3- a]quinoxalin-1-one (ODQ, of guanylyl cyclase), KT-5823 (of PKG activation), and iberiotoxin (IBX, of BKCa2+ activity) do not alter the [Ca2+]i response to KCl, suggesting no discernible endogenous NO production under basal conditions. The NO donor sodium nitroprusside (SNP) reduces the [Ca2+]i response to 77 nM; IBX restores the response to control values. These data show that activation of BKCa2+ in the presence of NO/cGMP provides a brake on KCl-induced [Ca2+]i responses. Experiments with the inhibitor of cyclic ADPR 8-bromo-cyclic ADPR (8-Br-cADPR) and SNP + downstream inhibitors of PKG and BKCa2+ suggest that NO inhibits ADPR cyclase in intact arterioles. When we pretreat afferent arterioles with 8-bromoguanosine 3′,5′-cyclic monophosphate (8-Br-cGMP; 10 μM), the response to KCl is 143 nM. However, in the presence of both IBX and 8-Br-cGMP, we observe a surprising doubling of the [Ca2+]i response to KCl. In summary, we present evidence for effects of the NO/cGMP/PKG system to reduce [Ca2+]i, via activation of BKCa2+ and possibly by inhibition of ADPR cyclase, and to increase [Ca2+]i, by a mechanism(s) yet to be defined.

scholarly journals Terminalia chebulaFructus Inhibits Migration and Proliferation of Vascular Smooth Muscle Cells and Production of Inflammatory Mediators in RAW 264.7

2015 ◽  
Vol 2015 ◽  
pp. 1-10 ◽  
Author(s):  
Hyun-Ho Lee ◽  
Keshav Raj Paudel ◽  
Dong-Wook Kim
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Inflammatory Mediators ◽  
Muscle Cells ◽  
No Production ◽  
Raw 264.7 ◽  
Terminalia Chebula

Pathogenesis of atherosclerosis and neointima formation after angioplasty involves vascular smooth muscle cells (VSMCs) migration and proliferation followed by inflammatory responses mediated by recruited macrophages in the neointima.Terminalia chebulais widely used traditional medicine in Asia for its beneficial effects against cancer, diabetes, and bacterial infection. The study was designed to determine whetherTerminalia chebulafructus water extract (TFW) suppresses VSMC migration and proliferation and inflammatory mediators production in macrophage (RAW 264.7). Our results showed that TFW possessed strong antioxidative effects in 1,1-diphenyl-2-picryl hydrazyl (DPPH) scavenging and lipid peroxidation assays. In addition, TFW reduced nitric oxide (NO) production, inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) expression in RAW 264.7 cells. Also, TFW inhibited platelet-derived growth factor (PDGF-BB) induced VSMC migration as determined by wound healing and Boyden chamber assays. The antimigratory effect of TFW was due to its inhibitory effect on metalloproteinase-9 (MMP-9) expression, focal adhesion kinase (FAK) activation, and Rho-family of small GTPases (Cdc42 and RhoA) expression in VSMCs. Furthermore, TFW suppressed PDGF-BB induced VSMC proliferation by downregulation of mitogen activated protein kinases (MAPKs) signaling molecules. These results suggest that TFW could be a beneficial resource in the prevention of atherosclerosis.

Estimation of nitric oxide production and reactionrates in tissue by use of a mathematical model

1998 ◽  
Vol 274 (6) ◽  
pp. H2163-H2176 ◽  
Author(s):  
Mark W. Vaughn ◽  
Lih Kuo ◽  
James C. Liao
Keyword(s):  
Nitric Oxide ◽  
Mathematical Model ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Consumption Rate ◽  
Physical Parameters ◽  
No Production ◽  
Rate Expression ◽  
Order Rate ◽  
Lack Of Information

Nitric oxide (NO) produced by the vascular endothelium is an important biologic messenger that regulates vessel tone and permeability and inhibits platelet adhesion and aggregation. NO exerts its control of vessel tone by interacting with guanylyl cyclase in the vascular smooth muscle to initiate a series of reactions that lead to vessel dilation. Previous efforts to investigate this interaction by mathematical modeling of NO diffusion and reaction have been hampered by the lack of information on the production and degradation rate of NO. We use a mathematical model and previously published experimental data to estimate the rate of NO production, 6.8 × 10−14μmol ⋅ μm−2 ⋅ s−1; the NO diffusion coefficient, 3,300 μm2s−1; and the NO consumption rate coefficient in the vascular smooth muscle, 0.01 s−1 (1st-order rate expression) or 0.05 μM−1 ⋅ s−1 (2nd-order rate expression). The modeling approach is discussed in detail. It provides a general framework for modeling the NO produced from the endothelium and for estimating relevant physical parameters.

Vascular smooth muscle cells on Matrigel as a model for LPS-induced hypocontractility and NO formation

1997 ◽  
Vol 272 (1) ◽  
pp. H576-H584 ◽  
Author(s):  
S. Li ◽  
S. X. Fan ◽  
T. M. McKenna
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
No Production ◽  
Cell Contractility ◽  
No Formation ◽  
Low Levels

Treatment of vascular tissue with low levels of lipopolysaccharide (LPS) induces nitric oxide synthase (NOS) activity and diminishes vascular contractility. However, in cultured vascular smooth muscle cells (VSMC), very high doses of LPS or the combination of LPS with cytokines are required for the induction of nitric oxide (NO) formation. The aims of this study were to establish a cell model to investigate LPS-induced hypocontractility and NO production and to test the hypothesis that responses of VSMC to LPS are differentiation regulated. We used Matrigel basement membrane matrix to maintain VSMC differentiation and found that VSMC cultured on Matrigel retained significant contractility in response to KCl stimulation. Incubation of VSMC with low levels of LPS(1–100 ng/ml) induced NOS mRNA and protein, induced NO production, and decreased cell contractility in a time- and dose-dependent fashion. The NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) partially restored LPS-treated VSMC contractility, whereas L-arginine reversed the contractility-restoring effect of L-NAME. These results suggest that VSMC grown on Matrigel are a useful experimental model for investigations into signal transduction mechanisms responsible for LPS-induced vascular hypocontractility.

The role of nitric oxide synthase/nitric oxide in vascular smooth muscle control

Perfusion ◽  
2000 ◽  
Vol 15 (2) ◽  
pp. 97-104 ◽  
Author(s):  
D Bradford Sanders ◽  
Tara Kelley ◽  
Douglas Larson
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Nitric Oxide Synthase ◽  
Vascular Smooth Muscle ◽  
Vascular Function ◽  
No Production ◽  
Amyl Nitrite ◽  
Vascular Compliance ◽  
Oxide Synthase

Vascular compliance is dependent on endogenous and exogenous sources of nitric oxide (NO). In a discussion of therapeutics and NO derived via nitric oxide synthase (NOS) enzymes, it is necessary to examine the pathways of each drug to provide the clinical perfusionist with a greater understanding of the role of NOS/NO in vascular function. Endothelial-derived NO is a contributor in the vasoregulation of vascular smooth muscle. Therapeutics seek to mimic the vasodilatory effects of the endogenous NO. The therapeutics included in this review are nitroglycerin, nitroprusside, amyl nitrite, and inhalation of NO. L-Arginine supplementation provides additional substrate for the endogenous pathway that can augment NO production. NO is a small bioactive molecule involved in various biochemical pathways. Dysregulation of NO production can impair normal physiologic control of vascular compliance. Therefore, the purpose of this review is to provide the perfusionist with an understanding of the biochemical and pharmacological aspects of NOS/NO associated with vascular function.

scholarly journals The Inhibitory Effect of Leptin on Angiotensin II-Induced Vasoconstriction in Vascular Smooth Muscle Cells Is Mediated via a Nitric Oxide-Dependent Mechanism

Endocrinology ◽  
2007 ◽  
Vol 148 (1) ◽  
pp. 324-331 ◽  
Author(s):  
Amaia Rodríguez ◽  
Ana Fortuño ◽  
Javier Gómez-Ambrosi ◽  
Guillermo Zalba ◽  
Javier Díez ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Smooth Muscle ◽  
Vascular Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Vascular Smooth Muscle Cells ◽  
Muscle Cells ◽  
No Production ◽  
Organ Bath ◽  
Ang Ii ◽  
Inos Protein

Leptin inhibits the contractile response induced by angiotensin (Ang) II in vascular smooth muscle cells (VSMCs) of the aorta. We studied in vitro and ex vivo the role of nitric oxide (NO) in the effect of leptin on the Ang II-induced vasoconstriction of the aorta of 10-wk-old Wistar rats. NO and nitric oxide synthase (NOS) activity were assessed by the Griess and 3H-arginine/citrulline conversion assays, respectively. Stimulation of inducible NOS (iNOS) as well as Janus kinases/signal transducers and activators of transcription (JAK/STAT) and phosphoinositide 3-kinase (PI3K)/Akt signaling pathways were determined by Western blot. The contractile responses to Ang II were evaluated in endothelium-denuded aortic rings using the organ bath system. Changes in intracellular Ca2+ were measured in VSMCs using fura-2 fluorescence. Leptin significantly (P ≤ 0.01) stimulated NO release and NOS activity in VSMCs. Leptin’s effect on NO was abolished by the NOS inhibitor, NG-monomethyl l-arginine, or the iNOS selective inhibitor l-N6-(1-iminoethyl)-lysine. Accordingly, leptin increased iNOS protein expression, with a comparable time course with that of NO production and NOS activity. Leptin also significantly increased STAT3 (P ≤ 0.01) and Akt (P ≤ 0.001) phosphorylation. Moreover, either the JAK2 inhibitor, AG490, or the PI3K inhibitor, wortmannin, significantly (P ≤ 0.05) abrogated the leptin-induced increase in iNOS protein. Finally, both NG-monomethyl l-arginine and l-N6-(1-iminoethyl)-lysine inhibitors completely blunted (P ≤ 0.001) the leptin-mediated inhibition of the Ang II-induced VSMC activation and vasoconstriction. These findings suggest that the endothelium-independent depressor action of leptin is mediated by an increase of NO bioavailability in VSMCs. This process requires the up-regulation of iNOS through mechanisms involving JAK2/STAT3 and PI3K/Akt pathways.

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