scholarly journals Differential regulation of nitric oxide synthase mRNA expression by lipopolysaccharide and pro-inflammatory cytokines in fetal hepatocytes treated with cycloheximide

1997 ◽  
Vol 327 (3) ◽  
pp. 819-823 ◽  
Author(s):  
Marta CASADO ◽  
María J. M DÍAZ-GUERRA ◽  
Lisardo BOSCÁ ◽  
Paloma MARTÍN-SANZ
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Mrna Expression ◽  
Inflammatory Cytokines ◽  
Inos Mrna ◽  
Mrna Levels ◽  
Mobility Shift ◽  
Fetal Hepatocytes ◽  
Oxide Synthase ◽  
Pro Inflammatory Cytokines

The effect of cycloheximide (CHX) on the mRNA expression of the cytokine-inducible, calcium-independent nitric oxide synthase (iNOS) was investigated in fetal hepatocytes stimulated with lipopolysaccharide (LPS) or pro-inflammatory cytokines. In the presence of CHX the LPS-dependent iNOS mRNA levels were reduced, whereas the response to pro-inflammatory cytokines was enhanced. Because iNOS transcription is highly dependent on the activation of nuclear factor κB (NF-κB), this factor was evaluated by electrophoretic mobility shift assays, and a close correlation between NF-κB activity and iNOS mRNA levels was observed. CHX itself potentiated the degradation of the IκBα and IκBβ inhibitory subunits (IκB is inhibitory κB) of the NF-κB complex, and therefore the loss of LPS-dependent iNOS mRNA expression cannot be attributed to a blockage in the activation of NF-κB. These results suggest the existence of a CHX-sensitive pathway in the expression of iNOS mediated by LPS, a mechanism that is not involved in the response to pro-inflammatory cytokines.

scholarly journals Dexamethasone differentially affects interleukin 1β- and cyclic AMP-induced nitric oxide synthase mRNA expression in renal mesangial cells

1994 ◽  
Vol 304 (2) ◽  
pp. 337-340 ◽  
Author(s):  
D Kunz ◽  
G Walker ◽  
J Pfeilschifter
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Cyclic Amp ◽  
Mesangial Cells ◽  
Inos Mrna ◽  
Mrna Levels ◽  
Necrosis Factor Alpha ◽  
Camp Analogue ◽  
Oxide Synthase ◽  
Inos Induction

Inducible nitric oxide synthase (iNOS) is expressed in renal mesangial cells in response to two principal classes of activating signals that interact in a synergistic fashion. These two groups of activators comprise inflammatory cytokines such as interleukin (IL)-1 beta or tumour necrosis factor alpha and agents that elevate cellular levels of cyclic AMP (cAMP). We examined whether dexamethasone differentially affects iNOS induction in response to IL-1 beta and a membrane-permeable cAMP analogue, N6,O-2′-dibutyryladenosine 3′,5′-phosphate (Bt2cAMP). Nanomolar concentrations of dexamethasone suppress IL-1 beta- as well as Bt2cAMP-induced iNOS protein expression and production of nitrite, the stable end product of nitric oxide (NO) formation. In contrast, dexamethasone prevents induction of iNOS mRNA in response to Bt2cAMP without affecting IL-1 beta-triggered increase in iNOS mRNA levels. These data suggest that dexamethasone acts at different levels, depending on the stimulus used to suppress iNOS induction in mesangial cells.

Distribution of CD14+ macrophages, CD4+, CD8+ lymphocytes and mRNA expression of inducible nitric oxide synthase in the endometrium of repeat breeding cows

2013 ◽  
Vol 16 (3) ◽  
pp. 443-451 ◽  
Author(s):  
W. Barański ◽  
J. Kaleczyc ◽  
S. Zduńczyk ◽  
W. Podlasz ◽  
E. Długołęcka-Malinowska ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Mrna Expression ◽  
Inducible Nitric Oxide Synthase ◽  
Polymorphonuclear Neutrophils ◽  
Control Group ◽  
Inos Mrna ◽  
Oxide Synthase ◽  
Subclinical Endometritis ◽  
Inducible Nitric Oxide

Abstract The expression of CD14+ macrophages, CD4+, CD8+ lymphocytes and mRNA of inducible nitric oxide synthase (iNOS) was investigated in the endometrium of repeat breeders with subclinical endometritis [experimental group (EXP), n = 10] and healthy [control group (CTRL), n = 10] cows. The cows were selected on the basis of repeat breeding (3 unsuccessful inseminations), clinical and cytological examinations (> 10% polymorphonuclear neutrophils in uterine smears obtained by cytobrush). From all the cows endometrial biopsies were collected and the presence of CD14+, CD4+ and CD8+ cells in the endometrium was evaluated immunohistochemically using semi quantitative counting method. The mRNA expression of iNOS was determined using reverse transcription-PCR. In general, there were no significant differences between EXP and CTRL groups in the expression of CD4+ and CD8 + lymphocytes in all endometrial structures. In contrast, we observed a higher number of CD14+ macrophages in repeat breeding group compared to the control cows, however, this difference was slightly pronounced. CD14+ cells were detectable only in the stratum compactum and stratum spongiosum. The statistically significant (p ≤ 0.05) higher expression of iNOS mRNA was measured in the cows with subclinical endometritis compared to the healthy animals. Our results suggest that the increased expression of CD14+ macrophages and iNOS mRNA may be associated with embryonal mortality in repeat breeding cows with subclinical endometritis.

Prolonged colonic epithelial hyporesponsiveness after colitis: role of inducible nitric oxide synthase

1999 ◽  
Vol 276 (3) ◽  
pp. G703-G710 ◽  
Author(s):  
Samuel Asfaha ◽  
Cameron J. Bell ◽  
John L. Wallace ◽  
Wallace K. MacNaughton
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Mrna Expression ◽  
Inducible Nitric Oxide Synthase ◽  
Mucosal Inflammation ◽  
Inos Mrna ◽  
Trinitrobenzenesulfonic Acid ◽  
Ussing Chambers ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

Colonic epithelial secretion is an important host defense mechanism. We examined whether a bout of colitis would produce long-lasting changes in epithelial function that persisted after resolution of mucosal inflammation. Colitis was induced in rats with intracolonic trinitrobenzenesulfonic acid. Six weeks later, colonic damage and inducible nitric oxide synthase (iNOS) mRNA expression and activity were measured. Segments of distal colon were mounted in Ussing chambers for measurement of permeability and responsiveness to secretory stimuli. Basal electrolyte transport parameters and permeability were not different from untreated controls. Despite normal macroscopic and histological appearance, secretory responses to electrical field stimulation (EFS), isobutylmethylxanthine (IBMX), and carbachol were significantly depressed (by 60–70%) relative to controls. iNOS mRNA expression and enzyme activity were significantly elevated. Dexamethasone reversed epithelial hyporesponsiveness and significantly reduced iNOS mRNA expression. A selective iNOS inhibitor normalized the secretory responses to EFS and IBMX but not to carbachol. These data suggest that ongoing synthesis of nitric oxide by iNOS contributes to chronic suppression of epithelial secretory function after episodes of colitis.

Inhibition of inducible nitric oxide synthesis by the herbal preparation Padma 28 in macrophage cell line

2000 ◽  
Vol 78 (11) ◽  
pp. 861-866 ◽  
Author(s):  
Thomas Moeslinger ◽  
Roswitha Friedl ◽  
Ivo Volf ◽  
Monika Brunner ◽  
Elisabeth Koller ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inducible Nitric Oxide Synthase ◽  
Inos Mrna ◽  
Mrna Levels ◽  
Nitric Oxide Synthesis ◽  
Mouse Macrophages ◽  
Oxide Synthase ◽  
Padma 28 ◽  
Inducible Nitric Oxide

Padma 28 is a mixture of herbs used in traditional Tibetan medicine with anti-inflammatory activities. We investigated the effects of Padma 28 on nitric oxide (NO) production by the inducible nitric oxide synthase (iNOS) in lipopolysaccharide stimulated mouse macrophages (RAW 264.7). Padma 28 (0-900 µg/mL) induced a concentration dependent inhibition of inducible nitric oxide synthesis. iNOS protein expression showed a concentration dependent reduction as revealed by immunoblotting when cells were incubated with increasing amounts of Padma 28. Padma 28 decreased iNOS mRNA levels as shown by RT-PCR. Aqueous extracts from costi amari radix (costus root, the dried root of Saussurea lappa) and the outer cover of myrobalani fructus (the dried fruit of Terminalia chebula), constituents of the complex herb preparation Padma 28, were found to inhibit inducible nitric oxide synthesis by decreasing iNOS protein and iNOS mRNA levels. The inhibition of inducible nitric oxide synthesis might contribute to the anti-inflammatory activities of Padma 28.Key words: inducible nitric oxide synthase, mouse macrophages, myrobalans, radix costae.

scholarly journals Expression of Endomyocardial Nitric Oxide Synthase and Coronary Endothelial Function in Human Cardiac Allografts

Circulation ◽  
2001 ◽  
Vol 104 (suppl_1) ◽  
Author(s):  
Stephen M. Wildhirt ◽  
Michael Weis ◽  
Costas Schulze ◽  
Nicole Conrad ◽  
Sinan Pehlivanli ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Endothelial Dysfunction ◽  
Nitric Oxide Synthase ◽  
Endothelial Function ◽  
Heart Transplantation ◽  
Mrna Expression ◽  
Inos Mrna ◽  
Inos Protein ◽  
Cardiac Allografts ◽  
Oxide Synthase

Background Inducible nitric oxide synthase (iNOS) is expressed and is functionally active in the presence of transplant arteriosclerosis. However, the early involvement of iNOS in alterations of microvascular endothelial function in the absence of preexisting lesions remains unclear; this information would be of prognostic value. We studied the course of iNOS mRNA expression, transcardiac nitric oxide production, and their potential association with microvascular coronary endothelial dysfunction in human cardiac allografts. Methods and Results A total of 42 patients were studied at 1, 6, and 12 months after heart transplantation. Microvascular coronary flow velocity reserve (CFVR) was tested in an endothelium-dependent (acetylcholine) and -independent manner (adenosine) using a Doppler flow wire. Endomyocardial iNOS expression was determined by reverse transcription polymerase chain reaction. iNOS protein and nitrotyrosine levels were detected by immunohistochemistry. Transcardiac plasma nitrite/nitrate (NOx) levels were measured by the Griess reaction. CFVR was impaired in 26.1% of patients (n=11) at 1 month and in 31% of patients (n=13) at 12 months after heart transplantation. Patients who developed impaired CFVR in the first year showed a significant increase in iNOS gene expression. Patients with impairment of CFVR 1 month after heart transplantation had higher levels of iNOS mRNA than patients with a normal CFVR. Patients with an initial impairment of CFVR who did not improve over time presented with significantly higher iNOS mRNA levels. iNOS protein and nitrotyrosine were expressed in the endomyocardial vessels of patients with impaired CFVR. Transcardiac NOx release was higher in patients with impaired CFVR. Conclusions In human cardiac allografts, microvascular endothelial dysfunction is associated with an enhanced endomyocardial iNOS mRNA expression and higher transcardiac NOx production and is accompanied by the expression of nitrotyrosine protein, suggesting peroxynitrite plays a role in the disease process. The data from the present study suggest an important role for the iNOS/nitric oxide pathway in the regulation of microvascular function in the absence of preexisting atherosclerotic lesions.

scholarly journals Pentoxifylline reduces the inflammatory process in diabetic rats: relationship with decreases of pro-inflammatory cytokines and inducible nitric oxide synthase

2015 ◽  
Vol 12 (1) ◽  
Author(s):  
Francisca Adilfa de Oliveira Garcia ◽  
Jéssica Farias Rebouças ◽  
Teresa Queiroz Balbino ◽  
Teresinha Gonçalves da Silva ◽  
Carlson Hélder Reis de Carvalho-Júnior ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Inflammatory Cytokines ◽  
Inducible Nitric Oxide Synthase ◽  
Inflammatory Process ◽  
Diabetic Rats ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide ◽  
Pro Inflammatory Cytokines
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