ANP and BNP Exert Anti-Inflammatory Action via NPR-1/cGMP Axis by Interfering with Canonical, Non-Canonical, and Alternative Routes of Inflammasome Activation in Human THP1 Cells

Dysregulated inflammasome activation and interleukin (IL)-1β production are associated with several inflammatory disorders. Three different routes can lead to inflammasome activation: a canonical two-step, a non-canonical Caspase-4/5- and Gasdermin D-dependent, and an alternative Caspase-8-mediated pathway. Natriuretic Peptides (NPs), Atrial Natriuretic Peptide (ANP) and B-type Natriuretic Peptide (BNP), binding to Natriuretic Peptide Receptor-1 (NPR-1), signal by increasing cGMP (cyclic guanosine monophosphate) levels that, in turn, stimulate cGMP-dependent protein kinase-I (PKG-I). We previously demonstrated that, by counteracting inflammasome activation, NPs inhibit IL-1β secretion. Here we aimed to decipher the molecular mechanism underlying NPs effects on THP-1 cells stimulated with lipopolysaccharide (LPS) + ATP. Involvement of cGMP and PKG-I were assessed pre-treating THP-1 cells with the membrane-permeable analogue, 8-Br-cGMP, and the specific inhibitor KT-5823, respectively. We found that NPs, by activating NPR-1/cGMP/PKG-I axis, lead to phosphorylation of NLRP3 at Ser295 and to inflammasome platform disassembly. Moreover, by increasing intracellular cGMP levels and activating phosphodiesterases, NPs interfere with both Gasdermin D and Caspase-8 cleavage, indicating that they disturb non-canonical and alternative routes of inflammasome activation. These results showed that ANP and BNP anti-inflammatory and immunomodulatory actions may involve the inhibition of all the known routes of inflammasome activation. Thus, NPs might be proposed for the treatment of the plethora of diseases caused by a dysregulated inflammasome activation.

Download Full-text

Increased Expression of the cGMP-Specific Phophodiesterase, PDE9A, in Sickle Cell Disease (SCD) Reticulocytes and Neutrophils, and Induction of Erythroid γ Globin Expression and Reduction of SCD Neutrophil Adhesion Following PDE9A Inhibition.

Blood ◽

10.1182/blood.v110.11.3397.3397 ◽

2007 ◽

Vol 110 (11) ◽

pp. 3397-3397

Author(s):

Camila B. Almeida ◽

Sara T.O. Saad ◽

Fernando F. Costa ◽

Nicola Conran

Keyword(s):

Specific Inhibitor ◽

K562 Cells ◽

Cell Types ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Gene Expressions ◽

Adhesive Properties ◽

Pharmacological Inhibition ◽

Intracellular Cgmp

Abstract Modulation of the levels of the NO second messenger, cyclic guanosine monophosphate (cGMP), has been identified as a possible therapeutic approach for SCD treatment. Up-regulation of cGMP-dependent signaling may induce γ-globin production in erythrocytic lineage cells and diminish the adhesive properties of leukocytes, the adhesion of which contributes to vaso-occlusion. Since the phosphodiesterase (PDE) enzymes, which degrade intracellular cyclic nucleotides, display differing cellular expressions, their inhibition can provide tissue-specific induction of cGMP. We studied the gene expressions of 3 types of cGMP-specific PDE (−1A, −5A, −9A) in the reticulocytes (ret) and neutrophils (neu) of healthy controls (AA), steady-state SCD patients (SS) and SCD patients on hydroxyurea therapy (SSHU; 20–30 mg/kg/day). Ret and neu were separated from whole blood before extracting mRNA, synthesizing cDNA and quantifying PDE gene expression by Real-Time PCR relative to β-actin and GAPDH expression. PDE1 was not found expressed in ret of any of the groups, but was found expressed in the neu of all three groups (≤14.2 rel expression). Intermediate PDE5 expression was observed in ret of all groups (≤16.8 rel express); PDE5 expression in neu was, however, almost undetectable in all groups (<0.3 rel. express). In contrast, high PDE9A expressions were found in ret of all groups with significantly higher expression (P<0.05, unpaired t test) in the ret of SS than SSHU and AA (68.3±10.4; 181.8±50.0; 54.8±22.2 rel express for AA, SS, SSHU, resp., n=6). Extremely high PDE9A expressions were also found in the neu of all groups, although PDE9A was significantly higher in SCD neu than in control neu (101.7±24.0; 722.8±102.2; 873.9±221.7 rel. express for AA, SS, SSHU, resp., n≥4, P<0.05 for SS/SSHU comp. AA). Comparison of PDE9A expression in diverse cell types that included T98G glial cells, liver, colon, skin, spleen, lymphnode, mammary, ovary, uterus, testicle and K562 erythroleukemic cells demonstrated very low level or virtually undetectable gene PDE9A expression in all of these tissue types (<6.7 rel express in all tissues), with the exception of K562 cells (96.8 rel. express). Data indicate that PDE9A may be expressed at higher levels in hematopoietic lineage cells. In keeping with our results, co-culture of K562 cells in the presence of the PDE1 and PDE5 inhibitors, vinpocetine (1–50μM) and sildenafil (50–200μg/ml), respectively, did not significantly increase intracellular cGMP, nor γ-globin expression (data not shown, P>0.05), suggesting that PDE1 and PDE5 are not suitable targets for pharmacological inhibition in erythroid lineage cells. In contrast, co-culture of K562 cells in the presence of the PDE9A-specific inhibitor, BAY 73-6691, for 48 h significantly increased intracellular γ-globin expression by 97.0±7.8% (1.0μM, n=3, P<0.05). Furthermore, in vitro static adhesion assays showed that co-incubation of SS neu with BAY 73-6691 reversed augmented SS neu adhesion to immobilized fibronectin (20μg/ml) to control neu adhesion levels (AA neu adhesion: 8.7±1.3%; SS neu adhesion: 12.6±1.4%, reduced to 7.2±1.3% with 1.0 μM BAY 73-6691, n=4, P<0.01 comp.basal SS adhesion). The PDE9A enzyme may constitute a relatively hematopoietic cell-specific target for pharmacological inhibition with a potential to induce γ-globin production and to inhibit cell adhesion mechanisms, with beneficial consequences for individuals with SCD.

Download Full-text

C-type natriuretic peptide stimulates function of the murine Sertoli cells via activation of the NPR-B/cGMP/PKG signaling pathway

Acta Biochimica Polonica ◽

10.18388/abp.2020_5442 ◽

2021 ◽

Author(s):

Yuejin Yu ◽

Yao Chen ◽

Chunlei Mei ◽

Na Li ◽

Kejia Wu ◽

...

Keyword(s):

Signaling Pathway ◽

Sertoli Cell ◽

Natriuretic Peptide ◽

Sertoli Cells ◽

Vascular Endothelial Cells ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Reproductive Process ◽

Intracellular Cgmp ◽

Important Regulator

C-type natriuretic peptide (CNP) is an important regulator of the male reproductive process. Our previous investigations showed that CNP can significantly stimulate the mRNA expression of androgen-binding protein (Abp) and transferrin (Trf) in the rat Sertoli cells, but the pathways responsible for this process remain to be elucidated. We predict that CNP binds the natriuretic peptide receptor B (NPR-B) to regulate expression of ABP and TRF through the intracellular cyclic guanosine monophosphate (cGMP) pathway. To address this question, in this study, we first confirmed the expression and localization of CNP and NPR-B in rat testes by immunohistochemistry and western blotting. Then, ELISA and real-time PCR were performed to investigate the signaling pathway of CNP in Sertoli cells in rat testes. Our results showed that CNP was mainly localized in the germ cells and Leydig cells, and its receptor, NPR-B, was mostly expressed in the Sertoli cells and vascular endothelial cells. CNP supplementation in the Sertoli cell medium was accompanied by an increase in the amount of intracellular cGMP and in the production of Abp and Trf mRNA, whereas inhibition of PKG with KT5823 led to a decrease in the expression of Abp and Trf mRNA. Moreover, Abp and Trf mRNA were no longer elevated when we used liposome-mediated RNA interference technology to silence the NPR-B gene in a mouse Sertoli cell line (TM4). These results suggest that CNP contributes to the regulation of ABP and TRF in the Sertoli cells through the NPR-B/cGMP/PKG signaling pathways.

Download Full-text

Natriuretic Peptides, Their Receptors, and Cyclic Guanosine Monophosphate-Dependent Signaling Functions

Endocrine Reviews ◽

10.1210/er.2005-0014 ◽

2005 ◽

Vol 27 (1) ◽

pp. 47-72 ◽

Cited By ~ 640

Author(s):

Lincoln R. Potter ◽

Sarah Abbey-Hosch ◽

Deborah M. Dickey

Keyword(s):

Natriuretic Peptide ◽

Natriuretic Peptides ◽

Bone Growth ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Long Bone ◽

Paracrine Factors ◽

Dependent Protein ◽

Membrane Spanning ◽

Inactivating Mutations

Natriuretic peptides are a family of structurally related but genetically distinct hormones/paracrine factors that regulate blood volume, blood pressure, ventricular hypertrophy, pulmonary hypertension, fat metabolism, and long bone growth. The mammalian members are atrial natriuretic peptide, B-type natriuretic peptide, C-type natriuretic peptide, and possibly osteocrin/musclin. Three single membrane-spanning natriuretic peptide receptors (NPRs) have been identified. Two, NPR-A/GC-A/NPR1 and NPR-B/GC-B/NPR2, are transmembrane guanylyl cyclases, enzymes that catalyze the synthesis of cGMP. One, NPR-C/NPR3, lacks intrinsic enzymatic activity and controls the local concentrations of natriuretic peptides through constitutive receptor-mediated internalization and degradation. Single allele-inactivating mutations in the promoter of human NPR-A are associated with hypertension and heart failure, whereas homozygous inactivating mutations in human NPR-B cause a form of short-limbed dwarfism known as acromesomelic dysplasia type Maroteaux. The physiological effects of natriuretic peptides are elicited through three classes of cGMP binding proteins: cGMP-dependent protein kinases, cGMP-regulated phosphodiesterases, and cyclic nucleotide-gated ion channels. In this comprehensive review, the structure, function, regulation, and biological consequences of natriuretic peptides and their associated signaling proteins are described.

Download Full-text

3′:5′-cyclic guanosine monophosphate (cGMP) potentiates the inositol 1,4,5-trisphosphate-evoked Ca2+ release in guinea-pig hepatocytes

Biochemical Journal ◽

10.1042/bj3180849 ◽

1996 ◽

Vol 318 (3) ◽

pp. 849-855 ◽

Cited By ~ 26

Author(s):

Gilles GUIHARD ◽

Laurent COMBETTES ◽

Thierry CAPIOD

Keyword(s):

Protein Kinase ◽

Guinea Pig ◽

Kinase Inhibitor ◽

Cell Voltage ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Intracellular Cgmp ◽

Inositol 1,4,5 Trisphosphate ◽

Dependent Protein ◽

Positive Effect

The effect of cGMP on noradrenaline-induced intracellular Ca2+ mobilization was investigated in whole-cell voltage-clamped guinea-pig hepatocytes. Treatment of the cells with 8-Br-cGMP (1–500 µM) resulted in an increase in the sensitivity of the cells to noradrenaline and to inositol 1,4,5-trisphosphate (InsP3) photoreleased from caged InsP3. The positive effect of 8-Br-cGMP on the Ca2+ release evoked by Ca2+-mobilizing agonists or InsP3 was blocked by a protein kinase G (PKG; cGMP-dependent protein kinase) inhibitor, the Rp-8-(4-chlorophenylthio)guanosine 3´:5´-monophosphorothioate. 8-Br-cGMP affected neither the basal InsP3 concentration nor the noradrenaline-induced production of InsP3. In permeabilized hepatocytes, the dose–response curve for InsP3-induced Ca2+ release was shifted to the left in the presence of 8-Br-cGMP. Furthermore, the treatment with 8-Br-cGMP did not affect the Ca2+ content of the InsP3-sensitive Ca2+ stores. These results indicate that intracellular cGMP potentiates the noradrenaline-induced Ca2+ response by enhancing Ca2+ release from the intracellular Ca2+ stores. We suggest that cGMP increases the apparent affinity of InsP3 receptors for InsP3 in guinea-pig hepatocytes probably by phosphorylation via the activation of PKG.

Download Full-text

The cGMP-Dependent Protein Kinase 2 Contributes to Cone Photoreceptor Degeneration in the Cnga3-Deficient Mouse Model of Achromatopsia

International Journal of Molecular Sciences ◽

10.3390/ijms22010052 ◽

2020 ◽

Vol 22 (1) ◽

pp. 52

Author(s):

Mirja Koch ◽

Constanze Scheel ◽

Hongwei Ma ◽

Fan Yang ◽

Michael Stadlmeier ◽

...

Keyword(s):

Protein Kinase ◽

Mouse Model ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Cone Photoreceptor ◽

Photoreceptor Degeneration ◽

Dependent Protein Kinase ◽

Genetic Ablation ◽

Cgmp Dependent Protein Kinase ◽

Dependent Protein

Mutations in the CNGA3 gene, which encodes the A subunit of the cyclic guanosine monophosphate (cGMP)-gated cation channel in cone photoreceptor outer segments, cause total colour blindness, also referred to as achromatopsia. Cones lacking this channel protein are non-functional, accumulate high levels of the second messenger cGMP and degenerate over time after induction of ER stress. The cell death mechanisms that lead to loss of affected cones are only partially understood. Here, we explored the disease mechanisms in the Cnga3 knockout (KO) mouse model of achromatopsia. We found that another important effector of cGMP, the cGMP-dependent protein kinase 2 (Prkg2) is crucially involved in cGMP cytotoxicity of cones in Cnga3 KO mice. Virus-mediated knockdown or genetic ablation of Prkg2 in Cnga3 KO mice counteracted degeneration and preserved the number of cones. Analysis of markers of endoplasmic reticulum stress and unfolded protein response confirmed that induction of these processes in Cnga3 KO cones also depends on Prkg2. In conclusion, we identified Prkg2 as a novel key mediator of cone photoreceptor degeneration in achromatopsia. Our data suggest that this cGMP mediator could be a novel pharmacological target for future neuroprotective therapies.

Download Full-text

Release of cyclic guanosine monophosphate evaluated as a diagnostic tool in cardiac diseases

Clinical Chemistry ◽

10.1093/clinchem/37.2.186 ◽

1991 ◽

Vol 37 (2) ◽

pp. 186-190 ◽

Cited By ~ 39

Author(s):

Karl-P Vorderwinkler ◽

Eilka Artner-Dworzak ◽

Gab Jakob ◽

Johanne Mair ◽

Franz Diensti ◽

...

Keyword(s):

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Cardiac Diseases ◽

Blood Samples ◽

Cutoff Value ◽

Intracellular Cgmp ◽

Glycerol Trinitrate ◽

Clinical Measurements ◽

And Storage

Abstract Concentrations of atrial natriuretic peptide (ANP) are increased in plasma of patients with impaired cardiac and renal function. The second messenger of ANP, cyclic guanosine monophosphate (cGMP), is released into the plasma specifically upon stimulation of cells with ANP. Although nitrates can also activate intracellular cGMP synthesis, we detected no increase in plasma cGMP concentrations after infusions of glycerol trinitrate. Because immunoreactive ANP is highly susceptible to degradation and nonspecific influences in blood samples, determinations of ANP require immediate centrifugation and storage of plasma at -20 degrees C. In contrast, we found that cGMP is stable for five days in vitro in blood samples containing EDTA. In 147 healthy blood donors, the upper cutoff value for plasma cGMP was 6.60 nmol/L, not significantly different (P greater than 0.05) from that for 222 patients with disorders other than cardiovascular and renal. In 69 patients with manifest congestive heart failure (NYHA stages II-IV), 65 had increased cGMP values. Using the above cutoff value for cGMP gave diagnostic sensitivity of 94.2% and specificity of 93.7%. Plasma cGMP may thus provide an alternative for routine clinical measurements of ANP in cardiac diseases in the absence of renal disorders.

Download Full-text

Effects of Atrial Natriuretic Peptide and Cyclic Guanosine Monophosphate on Isolated Human Myometrial Arteries Preconstricted by Endothelin-1

Gynecologic and Obstetric Investigation ◽

10.1159/000292333 ◽

1995 ◽

Vol 40 (3) ◽

pp. 190-194 ◽

Cited By ~ 7

Author(s):

Karolina-Rasa Kublickiene ◽

Charlotta Grunewald ◽

Marius Kublickas ◽

Bo Lindblom ◽

Nils-Olov Lunell ◽

...

Keyword(s):

Atrial Natriuretic Peptide ◽

Natriuretic Peptide ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Endothelin 1 ◽

Atrial Natriuretic

Download Full-text

Is cyclic guanosine monophosphate the internal 'second messenger' for cholinergic actions on central neurons?

Canadian Journal of Physiology and Pharmacology ◽

10.1139/y76-029 ◽

1976 ◽

Vol 54 (2) ◽

pp. 172-176 ◽

Cited By ~ 45

Author(s):

K. Krnjević ◽

E. Puil ◽

R. Werman

Keyword(s):

Time Course ◽

Membrane Conductance ◽

Cyclic Guanosine Monophosphate ◽

Guanosine Monophosphate ◽

Resting Potential ◽

Electrical Excitability ◽

Spinal Motoneurons ◽

Central Neurons ◽

Intracellular Cgmp ◽

Post Synaptic Potential

The most consistent effects produced by intracellular injections of guanosine 3′,5′-cyclic monophosphate (cGMP) (but not 5′-guanosine 5′-monophosphate in spinal motoneurons of cats are a rise in membrane conductance, acceleration in time course of spike potentials, and accentuation of the post-spike hyperpolarization. Associated changes in resting potential are smaller, less constant, and more often in the depolarizing than hyperpolarizing direction. cGMP tends to increase electrical excitability but reduces excitatory post-synaptic potential amplitudes. Most of the effects of intracellular cGMP are quite different from, or indeed opposite to, those of either extra- or intracellular applications of acetylcholine and therefore not consistent with the proposal that cGMP is the internal mediator of muscarinic actions.

Download Full-text