16K prolactin induces NF-kappaB activation in pulmonary fibroblasts

The amino-terminal 16 kDa fragment of prolactin (16K PRL) promotes the expression of the inducible isoform of nitric oxide synthase (iNOS) accompanied by the production of nitric oxide (NO) by rat pulmonary fibroblasts. The present study was designed to elucidate whether the mechanism by which 16K PRL promotes iNOS expression involves the activation of nuclear factor-kappa B (NF-kappaB), a key transcription factor for iNOS induction. 16K PRL stimulated DNA-binding activity of NF-kappaB in pulmonary fibroblasts as demonstrated by gel shift assays. Likewise, fluorescence immunocytochemistry showed that 16K PRL promotes nuclear translocation of the p65 subunit of NF-kappaB. Finally, treatment with 16K PRL induced the degradation of the NF-kappaB inhibitor kappaB-beta (IkappaB-beta), and such degradation was prevented by blocking IkappaB-beta phosphorylation. Altogether, these results show that 16K PRL activates NF-kappaB nuclear translocation via the phosphorylation and degradation of IkappaB-beta. These findings are consistent with NF-kappaB being part of the signal transduction pathway activated by 16K PRL to induce iNOS expression.

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Abstract 121: GRK5-NT Regulates the Activity of Calcium-Calmodulin-Dependent Transcription Factors

Circulation Research ◽

10.1161/res.111.suppl_1.a121 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Daniela Sorriento ◽

Anna Fusco ◽

Gaetano Santulli ◽

Bruno Trimarco ◽

Guido Iaccarino

Keyword(s):

Transcription Factors ◽

Cardiac Hypertrophy ◽

Nuclear Translocation ◽

Binding Activity ◽

Activated T Cells ◽

Calmodulin Binding ◽

Amino Terminal ◽

Calcium Calmodulin Dependent ◽

Dna Binding Activity

We have demonstrated that the amino terminal domain (GRK5-NT), which includes the RH domain and the region of binding to calmodulin, is able to regulate cardiac hypertrophy through inhibition of NFκB. Several studies indicate that mechanical stress and neuro-hormonal activation (Angiotensin, phenylephrine, endothelin) increase the levels of intracellular calcium, therefore activating calcineurin, a serine/threonine phosphatase that dephosphorylates nuclear factor of activated T cells (NFAT). Aim of this study is to evaluate the possibility that GRK5-NT regulates calcium-calmodulin dependent transcription factors. To this aim, we infected cardiomyoblasts in culture with an adenovirus encoding for GRK5-NT (AdGRK5-NT) or treated with a synthetic protein (TAT-RH), which reproduce the only RH domain of GRK5. Hypertrophic responses were induced by chronic stimulation of α 1 adrenergic receptors with phenylephrine (PE 10 -7 M,24h) and the levels of GATA4 and NFAT3 were assessed by western blot. PE induces activation and nuclear translocation of GATA4 and NFAT3 (NFAT:+38±3%;GATA4:+46±3% vs control), the treatment with AdGRK5-NT, but TAT-RH, reduces this effect (NFAT:-68±5%;GATA4:-56±2% vs PE). These data suggest that GRK5-NT using the NT domain regulates the activation of calcium-calmodulin dependent transcription factors through a mechanism of competition for binding to calmodulin. To confirm these data, we evaluated the effect of GRK5-NT in vivo in spontaneously hypertensive and hypertrophic rats (SHR). The overexpression of GRK5-NT in the heart was induced by intracardiac injection of 10 10 pfu/ml of AdGRK5-NT. The treatment inhibits nuclear translocation of NFAT3 and GATA4 (NFAT:-55±1%;GATA4:-34±3% vs PE) and is associated with a reduction in their DNA binding activity, as assessed by EMSA (NFAT:-47±1.5%;GATA4:-33±1% vs EP). In conclusion, our data indicate that GRK5-NT is able to regulate cardiac hypertrophy in two ways: inhibition of NFκB through binding and stabilization of IκB nuclear and inhibition activity of calcium- calmodulin dependent transcription factors, possibly by competing with calmodulin binding.

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Age alters cerebrovascular inflammation and effects of estrogen

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.01057.2006 ◽

2007 ◽

Vol 292 (5) ◽

pp. H2333-H2340 ◽

Cited By ~ 29

Author(s):

Lorraine Sunday ◽

Christa Osuna ◽

Diana N. Krause ◽

Sue P. Duckles

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Dna Binding ◽

Inducible Nitric Oxide Synthase ◽

Binding Activity ◽

Cerebral Vessels ◽

Cerebral Blood Vessels ◽

Dna Binding Activity ◽

Oxide Synthase ◽

Inducible Nitric Oxide

In young adult females, estrogen treatment suppresses the cerebrovascular inflammatory response; this is mediated in part via NF-κB, a key regulator of inflammatory genes. To examine whether age modifies effects of estrogen on vascular inflammation in the brain, female rats, 3 and 12 mo of age, were ovariectomized; half were treated with estrogen for 4 wk. Cerebral blood vessels were isolated from the animals at 4 and 13 mo of age. Inflammation was induced by LPS, either injected in vivo or incubated with isolated vessels ex vivo. Basal levels of cytoplasmic NF-κB were significantly higher in cerebral vessels of young rats, but the ratio of nuclear to cytoplasmic levels was greater in middle-aged animals. LPS exposure increased nuclear NF-κB DNA binding activity, protein levels of inducible nitric oxide synthase and cyclooxygenase-2, and production of nitric oxide and PGE2 in cerebral vessels. All effects of LPS were markedly greater in vessels from the older animals. Estrogen significantly inhibited the LPS-induced increase in NF-κB DNA binding activity in cerebral vessels from animals at both ages. In 4-mo-old rats, estrogen also significantly suppressed LPS induction of inducible nitric oxide synthase and cyclooxygenase-2 proteins, as well as production of nitric oxide and PGE2. In contrast, in 13-mo-old females, estrogen did not significantly affect these indexes of cerebrovascular inflammation. Thus the protective, anti-inflammatory effect of estrogen on cerebral blood vessels that is observed in young adults may be attenuated in aged animals, which exhibit a greater overall cerebrovascular response to inflammatory stimuli.

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Inhibitory Effect of Inflexinol on Nitric Oxide Generation and iNOS Expression via Inhibition of NF-κBActivation

Mediators of Inflammation ◽

10.1155/2007/93148 ◽

2007 ◽

Vol 2007 ◽

pp. 1-9 ◽

Cited By ~ 10

Author(s):

Jae Woong Lee ◽

Moon Soon Lee ◽

Tae Hun Kim ◽

Hwa Jeong Lee ◽

Seong Su Hong ◽

...

Keyword(s):

Nitric Oxide ◽

Inflammatory Diseases ◽

Inhibitory Effect ◽

Binding Activity ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dna Binding Activity ◽

Cox 2 ◽

Oxide Synthase ◽

Inducible Nitric Oxide

Inflexinol, anent-kaurane diterpenoid, was isolated from the leaves ofIsodon excisus. Many diterpenoids isolated from the genusIsodon(Labiatae) have antitumor and antiinflammatory activities. We investigated the antiinflammatory effect of inflexinol in RAW 264.7 cells and astrocytes. As a result, we found that inflexinol (1, 5, 10μM) suppressed the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) as well as the production of nitric oxide (NO) in LPS-stimulated RAW 264.7 cells and astrocytes. Consistent with the inhibitory effect on iNOS and COX-2 expression, inflexinol also inhibited transcriptional and DNA binding activity of NF-κBvia inhibition ofIκBdegradation as well as p50 and p65 translocation into nucleus. These results suggest that inflexinol inhibits iNOS and COX-2 expression through inhibition of NF-κBactivation, thereby inhibits generation of inflammatory mediators in RAW 264.7 cells and astrocytes, and may be useful for treatment of inflammatory diseases.

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Inhibition of IFN-γ-induced STAT1 activation by 15- deoxy-Δ12,14-prostaglandin J2

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00515.2002 ◽

2003 ◽

Vol 284 (5) ◽

pp. E883-E891 ◽

Cited By ~ 12

Author(s):

Sarah M. Weber ◽

Anna L. Scarim ◽

John A. Corbett

Keyword(s):

Binding Activity ◽

Inos Expression ◽

Cytokine Signaling ◽

Hsp70 Expression ◽

No Production ◽

Β Cells ◽

Inflammatory Gene Expression ◽

Dna Binding Activity ◽

Ifn Γ ◽

Oxide Synthase

The inhibitory actions of 15-deoxy-Δ12,14-prostaglandin J2 (PGJ2) on inflammatory gene expression have been attributed to the ability of this prostaglandin to inhibit the activation of NF-κB. In this study, we have identified an additional signaling pathway sensitive to inhibition by PGJ2. We show that PGJ2 inhibits interferon (IFN)-γ-stimulated phosphorylation and DNA-binding activity of STAT1. The inhibitory actions on STAT1 phosphorylation are first apparent after a 1- to 2-h incubation and are maximal after a 6-h incubation with PGJ2, and they correlate with the expression of heat shock protein (HSP)70 in islets. In previous studies, we have correlated the inhibitory actions of PGJ2 on inducible nitric oxide synthase (iNOS) expression and NF-κB activation in response to IL-1 with the increased expression of HSP70. Using overexpression and antisense depletion, we provide evidence that HSP70 does not mediate the inhibitory actions of PGJ2 on IL-1-induced NF-κB or IFN-γ-induced STAT1 activation or cytokine-stimulated iNOS expression by β-cells. Last, we show that the inhibitory actions of a short 6-h pulse with PGJ2 on IL-1 plus IFN-γ-stimulated iNOS expression and NO production by β-cells are persistent for extended periods (≤48 h). These findings suggest that PGJ2 inhibits multiple cytokine-signaling pathways (IL-1 and IFN-γ), that the inhibitory actions are persistent for extended periods, and that increased HSP70 expression correlates with, but does not appear to mediate, the inhibitory actions of PGJ2 on IL-1 and IFN-γ signaling in β-cells.

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Anti-Inflammatory Effect of Phytoncide in an Animal Model of Gastrointestinal Inflammation

Molecules ◽

10.3390/molecules26071895 ◽

2021 ◽

Vol 26 (7) ◽

pp. 1895

Author(s):

Azra Memon ◽

Bae Yong Kim ◽

Se-eun Kim ◽

Yuliya Pyao ◽

Yeong-Geun Lee ◽

...

Keyword(s):

Nitric Oxide ◽

Animal Model ◽

Gastric Ulcer ◽

Dextran Sulfate ◽

Dextran Sulfate Sodium ◽

Inos Expression ◽

Gastrointestinal Inflammation ◽

Anti Inflammatory ◽

Oxide Synthase ◽

Inflammatory Effect

Background: Phytoncide is known to have antimicrobial and anti-inflammatory properties. Purpose: This study was carried out to confirm the anti-inflammatory activity of two types of phytoncide extracts from pinecone waste. Methods: We made two types of animal models to evaluate the efficacy, an indomethacin-induced gastroenteritis rat model and a dextran sulfate sodium-induced colitis mouse model. Result: In the gastroenteritis experiment, the expression of induced-nitric oxide synthase (iNOS), a marker for inflammation, decreased in the phytoncide-supplemented groups, and gastric ulcer development was significantly inhibited (p < 0.05). In the colitis experiment, the shortening of the colon length and the iNOS expression were significantly suppressed in the phytoncide-supplemented group (p < 0.05). Conclusions: Through this study, we confirmed that phytoncide can directly inhibit inflammation in digestive organs. Although further research is needed, we conclude that phytoncide has potential anti-inflammatory properties in the digestive tract and can be developed as a functional agent.

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Isoquinolinequinone Derivatives from a Marine Sponge (Haliclona sp.) Regulate Inflammation in In Vitro System of Intestine

Marine Drugs ◽

10.3390/md19020090 ◽

2021 ◽

Vol 19 (2) ◽

pp. 90

Author(s):

Yun Kim ◽

Yeong Ji ◽

Na-Hyun Kim ◽

Nguyen Van Tu ◽

Jung-Rae Rho ◽

...

Keyword(s):

Nitric Oxide ◽

Marine Sponge ◽

Nuclear Translocation ◽

Hydroxyl Group ◽

Subsequent Increase ◽

Inflammatory Bowel ◽

Anti Inflammatory ◽

Key Functional Groups ◽

Oxide Synthase

Using bio-guided fractionation and based on the inhibitory activities of nitric oxide (NO) and prostaglandin E2 (PGE2), eight isoquinolinequinone derivatives (1–8) were isolated from the marine sponge Haliclona sp. Among these, methyl O-demethylrenierate (1) is a noble ester, whereas compounds 2 and 3 are new O-demethyl derivatives of known isoquinolinequinones. Compound 8 was assigned as a new 21-dehydroxyrenieramycin F. Anti-inflammatory activities of the isolated compounds were tested in a co-culture system of human epithelial Caco-2 and THP-1 macrophages. The isolated derivatives showed variable activities. O-demethyl renierone (5) showed the highest activity, while 3 and 7 showed moderate activities. These bioactive isoquinolinequinones inhibited lipopolysaccharide and interferon gamma-induced production of NO and PGE2. Expression of inducible nitric oxide synthase, cyclooxygenase-2, and the phosphorylation of MAPKs were down-regulated in response to the inhibition of NF-κB nuclear translocation. In addition, nuclear translocation was markedly promoted with a subsequent increase in the expression of HO-1. Structure-activity relationship studies showed that the hydroxyl group in 3 and 5, and the N-formyl group in 7 may be key functional groups responsible for their anti-inflammatory activities. These findings suggest the potential use of Haliclona sp. and its metabolites as pharmaceuticals treating inflammation-related diseases including inflammatory bowel disease.

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Activation of Nuclear Factor κB and Induction of Inducible Nitric Oxide Synthase by Ureaplasma urealyticumin Macrophages

Infection and Immunity ◽

10.1128/iai.68.12.7087-7093.2000 ◽

2000 ◽

Vol 68 (12) ◽

pp. 7087-7093 ◽

Cited By ~ 28

Author(s):

Y.-H. Li ◽

Z.-Q. Yan ◽

J. Skov Jensen ◽

K. Tullus ◽

A. Brauner

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Inducible Nitric Oxide Synthase ◽

Alveolar Macrophages ◽

Nuclear Factor Κb ◽

Inos Expression ◽

Dependent Manner ◽

Nuclear Factor ◽

Oxide Synthase ◽

Inducible Nitric Oxide

ABSTRACT Chronic lung disease (CLD) of prematurity is an inflammatory disease with a multifactorial etiology. The importance ofUreaplasma urealyticum in the development of CLD is debated, and steroids produce some improvement in neonates with this disease. In the present study, the capability of U. urealyticum to stimulate rat alveolar macrophages to produce nitric oxide (NO), express inducible nitric oxide synthase (iNOS), and activate nuclear factor κB (NF-κB) in vitro was characterized. The effect of NO on the growth of U. urealyticum was also investigated. In addition, the impact of dexamethasone and budesonide on these processes was examined. We found that U. urealyticum antigen (≥4 × 107 color-changing units/ml) stimulated alveolar macrophages to produce NO in a dose- and time-dependent manner (P < 0.05). This effect was further enhanced by gamma interferon (100 IU/ml; P < 0.05) but was attenuated by budesonide and dexamethasone (10−4 to 10−6 M) (P < 0.05). The mRNA and protein levels of iNOS were also induced in response to U. urealyticum and inhibited by steroids.U. urealyticum antigen triggered NF-κB activation, a possible mechanism for the induced iNOS expression, which also was inhibited by steroids. NO induced by U. urealyticum caused a sixfold reduction of its own growth after infection for 10 h. Our findings imply that U. urealyticum may be an important factor in the development of CLD. The host defense response againstU. urealyticum infection may also be influenced by NO. The down-regulatory effect of steroids on NF-κB activation, iNOS expression, and NO production might partly explain the beneficial effect of steroids in neonates with CLD.

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Abstract 316: Effects of Inhibiting of Inducible Nitric Oxide Synthase in the Pathophysiology of Experimental Preeclampsia

Hypertension ◽

10.1161/hyp.60.suppl_1.a316 ◽

2012 ◽

Vol 60 (suppl_1) ◽

Author(s):

Lorena M Amaral ◽

Ana Carolina T Palei ◽

Lucas C Pinheiro ◽

Jonas T Sertorio ◽

Danielle A Guimaraes ◽

...

Keyword(s):

Oxidative Stress ◽

Nitric Oxide ◽

Reactive Oxygen Species ◽

Mean Arterial Pressure ◽

Arterial Pressure ◽

Inducible Nitric Oxide Synthase ◽

Inos Expression ◽

Oxygen Species ◽

Oxide Synthase ◽

Inducible Nitric Oxide

The pathophysiology of preeclampsia (PE) is not entirely known. However, increased oxidative stress possibly leading to impaired nitric oxide activity has been implicated in the critical condition. Increased oxidative stress with increased levels of highly reactive species including superoxide may generate peroxynitrite. We examined the role of inducible nitric oxide synthase (iNOS) and oxidative stress in the reduced uterine perfusion pressure (RUPP) preeclampsia experimental model. METHODS: RUPP was induced in wistar rats. Pregnant rats in the RUPP group had their aortic artery clipped at day 14 of gestation. After a midline incision, a silver clip (0.203 mm) was placed around the aorta above the iliac bifurcation; silver clips (0.100 mm) were also placed on branches of both the right and left ovarian arteries that supply the uterus. Sham-operated (pregnant control rats) and RUPP rats were treated with oral vehicle or 1 mg/kg/day 1400W (iNOS inhibitor) for 5 days. Mean arterial pressure (MAP) and plasma levels of thiobarbituric acid-reactive species (TBARS) and total radical-trapping antioxidant potential (TRAP) were measured determined. Aortic iNOS expression (Western blotting) and reactive oxygen species (ROS; assessed by fluorescence microscopy with dihydroethidium-DHE) were measured. We found increased mean arterial pressure in RUPP compared with pregnant control rats (MAP= 128±1 vs. 100±1.8 mmHg, respectively; P<0.05) and 1400W exerted antihypertensive effects (MAP= 114±2 vs.128±1 mmHg in RUPP treated and untreated rats, respectively; P<0.05). Higher reactive oxygen species (ROS) concentrations were found in RUPP compared with pregnant control rats (7.1±0.5 vs. 5.1±0.5 arbitrary units (A.U.), respectively; P<0.05) and 1400W decreased ROS production to 5.8±0.02 A.U. in RUPP treated rats, P<0.05. In addition, 1400W attenuated iNOS expression in RUPP rats (0.29±0.02 vs. 0.55±0.8 A.U. in RUPP treated and untreated rats, respectively; P<0.01) and had no effects on plasma TBARS and TRAP levels. Our results suggest that 1400w exerts antihypertensive effects in the RUPP model and suppresses ROS formation. Supported by FAPESP,Cnpq.

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