Differential relaxing responses to particulate or soluble guanylyl  cyclase activation on endothelial cells: a mechanism dependent on PKG-Iα  activation by NO/cGMP

Rivero-Vilches, Francisco Javier; Frutos, Sergio de; Saura, Marta; Rodríguez‐Puyol, Diego; Rodrı́guez-Puyol, Manuel

doi:10.1152/ajpcell.00590.2002

Cited by 41 publications

(31 citation statements)

References 38 publications

(41 reference statements)

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“…10). This suggested compartmentalization of cGMP signaling, an emerging theme in multiple cell types (14,15,30,31,36). The observation that 8-pCPT-cGMP did not mimic the effect of pGC activation suggested that a cGMP-sensitive PDE was again involved rather than PKG I .…”

Section: Discussionmentioning

confidence: 99%

Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice

Schmidt

Damarla²,

Rentsendorj³

et al. 2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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(HV T) ventilation causes pulmonary endothelial barrier dysfunction. HV T ventilation also increases lung nitric oxide (NO) and cGMP. NO contributes to HV T lung injury, but the role of cGMP is unknown. In the current study, ventilation of isolated C57BL/6 mouse lungs increased perfusate cGMP as a function of V T. Ventilation with 20 ml/kg V T for 80 min increased the filtration coefficient (K f), an index of vascular permeability. The increased cGMP and Kf caused by HVT were attenuated by nitric oxide synthase (NOS) inhibition and in lungs from endothelial NOS knockout mice. Inhibition of soluble guanylyl cyclase (sGC) in wild-type lungs (10 M ODQ) also blocked cGMP generation and inhibited the increase in K f, suggesting an injurious role for sGC-derived cGMP. sGC inhibition also attenuated lung Evans blue dye albumin extravasation and wetto-dry weight ratio in an anesthetized mouse model of HV T injury. Additional activation of sGC (1.5 M BAY 41-2272) in isolated lungs at 40 min increased cGMP production and K f in lungs ventilated with 15 ml/kg V T. HVT endothelial barrier dysfunction was attenuated with a nonspecific phosphodiesterase (PDE) inhibitor (100 M IBMX) as well as an inhibitor (10 M BAY 60-7550) specific for the cGMPstimulated PDE2A. Concordantly, we found a V T-dependent increase in lung cAMP hydrolytic activity and PDE2A protein expression with a decrease in lung cAMP concentration that was blocked by BAY 60-7550. We conclude that HV T-induced endothelial barrier dysfunction resulted from a simultaneous increase in NO/sGC-derived cGMP and PDE2A expression causing decreased cAMP. nitric oxide; guanosine 3Ј,5Ј-cyclic monophosphate; phosphodiesterase 2A; adenosine 3Ј,5Ј-cyclic monophosphate; atrial natriuretic peptide; endothelial permeability MECHANICAL VENTILATION is necessary to support patients with the acute respiratory distress syndrome. However, mechanical ventilation also contributes to lung injury by a mechanism that directly correlates with tidal volume (V T ) (39). High V T (HV T ) ventilation causes pulmonary endothelial barrier dysfunction and edema, leading to ventilator-induced lung injury (VILI) (42). The mechanisms behind this injury are complex, but it is evident that HV T -induced changes in intracellular signaling play a significant role (12).Recent evidence from animal models of VILI implicates nitric oxide (NO) in the pathogenesis of HV T endothelial barrier dysfunction (4,6,29). NO is synthesized by three isoforms of nitric oxide synthase (NOS): endothelial NOS (eNOS), inducible NOS (iNOS), and neuronal NOS. Ventilatory lung stretch stimulates pulmonary microvascular NO production by an increase in eNOS expression (6), phosphorylation of eNOS by phosphatidylinositol 3-kinase-induced protein kinase B activation (22), or induction of iNOS expression (13,29). Once produced by NOS, NO reacts with reactive oxygen species (ROS) to generate peroxynitrite, a toxic free radical that contributes to VILI (29). NO can also stimulate endothelial soluble guanylyl cyclase (sGC) to generat...

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Section: Discussionmentioning

confidence: 99%

Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice

Schmidt

Damarla²,

Rentsendorj³

et al. 2008

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Scoparone also elevated the basal level of cGMP, which results in decreased phosphorylation of myosin light chain, then dilatation. 40,41) NO produced by the eNOS activation can dilate the vessel wall and scoparone may potentiate this vasodilative effect of NO. Paradoxically, scoparone has an inhibitory effect on NO generation through iNOS activation under our experimental conditions.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of Cytokine-Mediated Nitric Oxide Synthase Expression in Rat Insulinoma Cells by Scoparone

Kim

Kwon²,

Lee

et al. 2007

Biological & Pharmaceutical Bulletin

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Autoimmune diabetes is characterized by a local inflammatory reaction in and around the pancreatic islets, followed by selective destruction of insulin-producing b-cells.1) Much evidence supports an effector role for cytokines in mediating b-cell destruction.2,3) Incubation of rat islets with interleukin1b (IL-1b) alone, or in combination with tumor necrosis factor (TNF)-a and interferon-g (IFN-g) results in the expression of the inducible form of nitric oxide synthase (iNOS), increased production of nitric oxide (NO), and potent inhibition of glucose-stimulated insulin secretion. [4][5][6] Heitmeier et al. 7) demonstrated that IL-1b-induced inhibition of insulin secretion and nitrite production by rat islets were completely prevented by N w -nitro-L-arginine methylester (L-NAME) and aminoguanidine.One cellular target activated in response to IL-1b is the transcriptional regulator nuclear factor kB (NF-kB).8,9) NFkB is normally bound to inhibitory kB (IkB) in the cytosol; this binding prevents its movement into the nucleus. Various stimuli induce the phosphorylation of IkB, which releases NF-kB, and translocates to the nucleus, where it interacts with its DNA recognition sites to mediate gene transcription. 10,11) Scoparone (6,7-dimethoxycoumarin), a derivative of coumarin (1,2-benzopyrone) is known to have potent immunosuppressive, vascular relaxant and lipid lowering properties. [12][13][14][15] In this study, we examined the feasibility of scoparone as a means of preventing IL-1b and IFN-g-induced b-cell destruction. Scoparone inhibited IL-1b and IFN-g-induced NF-kB activation, iNOS expression, NO formation, glucose-stimulated insulin secretion (GSIS), and cell death of RIN cells, which may explain the beneficial effects of scoparone as an anti-diabetic agent. MATERIALS AND METHODSCell Culture RIN, clone 5F (RINm5F), is an insulinoma cell line derived from the NEDH rat islet cell tumor.16) Cells were purchased from the American Type Culture Collection and grown at 37°C under a humidified, 5% CO 2 atmosphere in RPMI 1640 (Gibco BRL) supplemented with 10% fetal bovine serum, 2 mM glutamine, 10000 units/ml of penicillin, 10 mg/ml of streptomycin, and 2.5 mg/ml of amphoptericin B.Cell Viability Assay The viability of cultured cells was determined by assaying the reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to formazan as described previously. 17) After treatment, cells in 96-well plates were washed twice with PBS. MTT (100 mg/100 ml of PBS) was added to each well. Cells were then incubated at 37°C for 1 h, and DMSO (100 ml) was added to dissolve the formazan crystals. Absorbance was measured at 570 nm with a model Spectra MAX PLUS (Molecular Devices).Cell Proliferation Assay A cell proliferation enzymelinked immunosorbent assay, BrdU kit (Amersham Biosciences, U.K.) was used to measure the incorporation of 5-bromo-2-deoxyuridine (BrdU) during DNA synthesis according to the manufacturer's protocol. Briefly, cells were seeded overnight in black 96-well tissue culture plates with c...

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“…Cyclic GMP was extracted from BAEC following published methods 26 ; cGMP production in each sample was measured by radioimmunoassay and corrected by protein amount (please see Supplemental data).…”

Section: Measurement Of Cyclic Gmpmentioning

confidence: 99%

Nitric Oxide Decreases the Expression of Endothelin-Converting Enzyme-1 Through mRNA Destabilization

Raoch

Rodrı́guez-Pascual²,

López-Martínez³

et al. 2011

ATVB

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Objective-Endothelial function depends on the equilibrium in the synthesis of vasoactive endothelial factors. It is well known that endothelin and nitric oxide (NO) exhibit reciprocal regulation. We assessed the ability of NO to regulate endothelin-converting enzyme-1 (ECE-1) expression in vascular endothelial cells. Methods and Results-Bovine aortic endothelial cells were incubated with 2 different NO donors as well as with a cyclic-GMP analog, dibutyryl-cGMP (dB-cGMP). ECE-1 protein content and mRNA expression were evaluated by Western blot and Northern blot, respectively, promoter activity by transfection experiments, ECE-1 activity by ELISA, and cGMP production by radioimmunoassay. Both NO donors decreased ECE-1 protein content, mRNA expression, and ECE-1 activity. ODQ, an inhibitor of soluble guanylyl cyclase, blocked those effects. NO donors raised cGMP levels, and dB-cGMP mimicked their effects on ECE-1 expression, which were blocked by KT5823, a nonspecific PKG inhibitor. The changes on ECE-1 expression were due to a destabilization on 3Ј-untranslated region (3Ј-UTR) of this mRNA, because the activity of a luciferase reporter construct containing the 3Ј-UTR of the ECE-1 gene was reduced by dB-cGMP in a PKG-dependent manner. The biological relevance of this regulation was confirmed in bovine aortic endothelial cells coincubated with macrophages in the presence of lipopolysaccharide, in eNOS-deficient mice, and in Wistar rats treated with NO donors. In every case, an inverse relationship was observed between NO and ECE-1 protein content. Conclusion-Our

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Differential relaxing responses to particulate or soluble guanylyl cyclase activation on endothelial cells: a mechanism dependent on PKG-Iα activation by NO/cGMP

Cited by 41 publications

References 38 publications

Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice

Soluble guanylyl cyclase contributes to ventilator-induced lung injury in mice

Inhibition of Cytokine-Mediated Nitric Oxide Synthase Expression in Rat Insulinoma Cells by Scoparone

Nitric Oxide Decreases the Expression of Endothelin-Converting Enzyme-1 Through mRNA Destabilization

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