scholarly journals Expression patterns of cytokines, p53 and nitric oxide synthase isoenzymes in corpora lutea of pseudopregnant rabbits during spontaneous luteolysis

Reproduction ◽  
2004 ◽  
Vol 127 (2) ◽  
pp. 229-238 ◽  
Author(s):  
Cristiano Boiti ◽  
Gabriella Guelfi ◽  
Massimo Zerani ◽  
Danilo Zampini ◽  
Gabriele Brecchia ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Protective Action ◽  
Expression Patterns ◽  
Physiological Role ◽  
P53 Gene ◽  
Total Activity ◽  
Inos Mrna ◽  
Corpora Lutea ◽  
Gene Expressions ◽  
Age Related

The gene expressions for macrophage chemoattractant protein-1 (MCP-1), interleukin (IL)-1β, IL-2 and p53 were examined by semi-quantitative RT-PCR in corpora lutea (CL) of rabbits during spontaneous luteolysis at days 13, 15, 18 and 22 of pseudopregnancy. In the same luteal tissue, total activity of nitric oxide (NO) synthase (NOS) and genes for both endothelial (eNOS) and inducible (iNOS) isoforms were also analysed. From day 13 to 15, MCP-1 and IL-1βmRNA levels rose (P≤ 0.01) almost 2-fold, and the transcript for p53 almost 8-fold, but then all dropped (P≤ 0.05) from day 18 onward. IL-2 mRNA abundance was higher (P≤ 0.01) on day 13 and then gradually declined. During luteolysis, eNOS mRNA decreased 40% (P≤ 0.05) by day 15, but thereafter remained unchanged, while iNOS mRNA was barely detectable and did not show any clear age-related pattern throughout the late luteal stages. Total NOS activity progressively increased (P≤ 0.01) from day 13 to 18 of pseudopregnancy and then dropped to the lowest (P≤ 0.01) levels on day 22. Luteal progesterone content also declined during CL regression from 411 to 17 pg/mg found on days 13 and 22 respectively, in parallel with the decrease in blood progesterone concentrations. These data further support a physiological role of NO as modulator of luteal demise in rabbits. Locally, luteal cytokines may be involved in the up-regulation of NOS activity, while downstream NO may inhibit steroroidogenesis and induce expression of p53 gene after removal of the protective action of progesterone.

scholarly journals Expression patterns of endothelial and inducible nitric oxide synthase isoforms in corpora lutea of pseudopregnant rabbits at different luteal stages

2002 ◽  
Vol 173 (2) ◽  
pp. 285-296 ◽  
Author(s):  
C Boiti ◽  
D Zampini ◽  
G Guelfi ◽  
F Paolocci ◽  
M Zerani ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Expression Patterns ◽  
Physiological Role ◽  
Total Activity ◽  
Pcr Analysis ◽  
Inos Mrna ◽  
Corpora Lutea ◽  
Mrna Levels ◽  
Isolated Cells ◽  
Protein Levels

Total activity of nitric oxide (NO) synthase (NOS) and expression of both endothelial (eNOS) and inducible (iNOS) isoforms were examined in corpora lutea (CL) of rabbits across pseudopregnancy by quantitative RT-PCR analysis, Western blot and immunohistochemistry. CL were collected at early- (day 4), mid- (day 9) and late- (day 13) luteal phases of pseudopregnancy. The PCR product of rabbit luteal eNOS was cloned and its direct sequence exhibited 90% homology with those of other species. The steady-state mRNA levels encoding eNOS remained fairly constant throughout both early- and mid-luteal stages of pseudopregnancy but dropped almost to half (P</=0.05) by day 13. By contrast, luteal eNOS proteins increased 2-fold (P</=0.05) from the early- to late-luteal phase. Independently of CL age, iNOS mRNA was very poorly expressed while protein levels gradually declined from the early- to late-luteal stage. Intense eNOS-like immunoreactivity was detected in large luteal cells, while iNOS staining was targeted to a few, isolated cells, probably macrophages. Basal NOS activity was greater in day 4 CL than in both day 9 and day 13 CL. These data are the first to characterize in rabbit CL the temporal expression patterns of NOS isoforms across different luteal stages of pseudopregnancy and, collectively, suggest the existence of an expressional control for this constitutive isoform, which might have a physiological role in regulating CL function during development.

scholarly journals Regulation of nitric oxide synthase isoforms and role of nitric oxide during prostaglandin F2alpha-induced luteolysis in rabbits

Reproduction ◽  
2003 ◽  
pp. 807-816 ◽  
Author(s):  
C Boiti ◽  
G Guelfi ◽  
D Zampini ◽  
G Brecchia ◽  
A Gobbetti ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Time Course ◽  
Plasma Concentrations ◽  
Physiological Role ◽  
Total Activity ◽  
Corpora Lutea ◽  
Prostaglandin F ◽  
Oxide Synthase

Total activity of nitric oxide synthase (NOS) and the gene expression of both endothelial NOS (eNOS) and inducible NOS (iNOS) isoforms in corpora lutea of pseudopregnant rabbits were examined during prostaglandin F(2alpha) (PGF(2alpha))-induced luteolysis. Corpora lutea were collected at 0, 6, 12, 24 and 48 h after an injection of PGF(2alpha) at day 9 of pseudopregnancy. At 12 h after PGF(2alpha) administration, luteal mRNA encoding eNOS decreased (P0.05) by 40% and remained low throughout the subsequent 36 h, whereas eNOS protein increased (P0.05) two- to threefold. By contrast, expression of mRNA encoding iNOS was poor and remained fairly constant, but transcription increased eightfold (P0.01) within 6 h after PGF(2alpha) treatment and then decreased to values similar to those of controls. Total NOS activity increased twofold (P0.01) at 6 h after treatment and remained high thereafter, whereas progesterone concentrations in explanted corpora lutea decreased (P0.01) from 302.4+/-42.3 pg x mg(-1) at day 9 to 58.6+/-8.3 at 48 h later, and peripheral plasma concentrations of progesterone declined too. Long-term administration of Nomega-nitro-L-arginine methyl ester (0.6 g l(-1) per os) from day 2 of pseudopregnancy onward partially blocked the luteolytic action of PGF(2alpha) administered at day 9 of pseudopregnancy. In nitric oxide (NO)-deficient rabbits, progesterone concentrations remained higher (P0.01) than in controls at 24-48 h after PGF(2alpha) administration (4.5 to 3.2 ng x ml(-1), respectively). These data are the first to characterize NOS activity. The time course of expression of eNOS and iNOS in rabbit corpora lutea during PGF(2alpha)-induced luteolysis gives additional support to a physiological role of NO in the regulation of regression of corpora lutea in rabbits.

scholarly journals Interleukin-1β and inducible form of nitric oxide synthase expression in early syngeneic islet transplantation

2007 ◽  
Vol 192 (1) ◽  
pp. 169-177 ◽  
Author(s):  
Marta Montolio ◽  
Montse Biarnés ◽  
Noèlia Téllez ◽  
Jessica Escoriza ◽  
Joan Soler ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cell Death ◽  
Nitric Oxide Synthase ◽  
Islet Transplantation ◽  
Interleukin 1Β ◽  
Inos Mrna ◽  
No Production ◽  
Gene Expressions ◽  
Syngeneic Islet Transplantation ◽  
Oxide Synthase

Islets are particularly vulnerable in the initial days after transplantation when cell death results in the loss of more than half of the transplanted islet tissue. To determine whether a non-specific inflammation at the grafted site mediated by the local expression of inflammatory cytokines could play a role on the initial damage to transplanted islets, we studied the expressions of interleukin-1β (IL-1β) and inducible form of nitric oxide synthase (iNOS) after syngeneic islet transplantation. Insulin-treated streptozotocin-diabetic Lewis rats were syngeneically transplanted with 500 islets. Grafts were harvested 1, 3, or 7 days after transplantation, and the expressions of IL-1β and iNOS genes were determined by RT-PCR. IL-1β and iNOS mRNAs were detected in islets immediately after isolation, and were upregulated after transplantation. IL-1β mRNA was ninefold increased on day 1, was still sevenfold increased on day 3 after transplantation, and declined towards pretransplantation levels on day 7. iNOS mRNA showed a similar pattern of expression to that of IL-1β: on days 1 and 3 after transplantation it was 14-and 4-fold higher respectively than in freshly isolated islets. In addition, IL-1β and iNOS were identified in islet grafts and found to be produced mainly by CD68-positive macrophages. A low number of IL-1β- and iNOS-positive but CD68-negative cells were also identified suggesting that other cell types, in addition to macrophages, were involved in the expression of IL-1β and NO production in islet grafts. The finding of increased IL-1β and iNOS gene expressions in the initial days after islet transplantation and the presence of IL-β and iNOS proteins in the graft confirmed the presence of an early non-specific inflammatory response after islet transplantation. Overall, the data suggest that IL-1β plays a role in the extensive β-cell death found in the initial days after islet transplantation.

scholarly journals Evaluation of the association between exosomal levels and female reproductive system and fertility outcome during aging: a systematic review protocol

2019 ◽  
Vol 8 (1) ◽  
Author(s):  
Halimeh Mobarak ◽  
Reza Rahbarghazi ◽  
Francesca Lolicato ◽  
Mohammad Heidarpour ◽  
Fariba Pashazadeh ◽  
...  
Keyword(s):  
Systematic Review ◽  
Reproductive System ◽  
Reproductive Tract ◽  
Physiological Role ◽  
Current Evidence ◽  
Gene Expressions ◽  
Younger Women ◽  
Age Related ◽  
Maternal Aging

Abstract Background Exosomes may have critical roles in the maternal-embryo cross-talk for the recognition and maintenance of pregnancy during maternal aging. Exosomes have the capability to carry developmental signaling molecules with the ability to modulate gene expressions and affect growth and regulation of embryo during pregnancy. Systematic review aims to evaluate age-related alterations in the exosomal content and functions that can influence the reproductive performance in human and animal models as conveyors of senescence signals. Methods A literature search of electronic databases including MEDLINE (PubMed), Embase, ProQuest, Scopus, Google Scholar, WHO, SID, MAGIRAN, and Barakat will be conducted. Following the online search, articles will be screened by two independent reviewers according to inclusion and exclusion criteria. Eligible studies will be critically appraised by reviewers at the study level for methodological quality using Joanna Briggs Institute’s standardized critical appraisal tools. The extracted data from selected studies will cover the study populations, methods, current evidence about the physiological role of extracellular vesicles and exosomes in reproductive system, relevant outcomes, and possible conclusions about the effectiveness of exposure. Discussion Regarding the role of exosomes and their cargoes in the function of reproductive tract, the possible beneficial or adverse effects following exosomal administration from younger women to older women will be evaluated in the systematic review. As a result, exosome therapy could be suggested as a novel therapeutic agent if the favorable results are identified.

Abstract 103: Interferon Regulatory Factor-1 Is Responsible For Cardiac Protective Effect Of 5-azacytidine On Macrophages In Myocardial Infarction

2014 ◽  
Vol 115 (suppl_1) ◽  
Author(s):  
Yong Sook Kim ◽  
Hye-yun Jeong ◽  
Youngkeun Ahn
Keyword(s):  
Nitric Oxide ◽  
Myocardial Infarction ◽  
Cardiac Function ◽  
Animal Study ◽  
Cardiac Fibrosis ◽  
Expression Patterns ◽  
Interferon Regulatory Factor ◽  
Inos Mrna ◽  
Regulatory Factor ◽  
Interferon Regulatory Factor 1

Background: During progression of cardiac injury, macrophages with specialized phenotypes are actively involved in inflammatory responses. In this study, we investigated the effect of 5-azacytidine (5AZ) on macrophages in the damaged myocardium. Methods: A mouse macrophage cell line RAW264.7 was stimulated with lipopolysaccharide (LPS, 100 ng/mL) with or without 5AZ (10 μM). Nitric oxide was quantified by Griess’s method. Expression patterns of inducible nitric oxide synthase (iNOS) and interferon regulatory factor 1 (IRF-1) were determined by Western blot analysis. For animal study, myocardial infarction (MI) was induced by ligation of left coronary artery in mice, and divided into four groups; non MI + saline, non-MI + 5AZ , MI + saline, and MI + 5AZ. Saline or 5AZ was injected (5 mg/kg/d) every other day. Cardiac fibrosis was evaluated by Masson’s trichrome stain was performed and cardiac function was by echocardiography 2 weeks after MI. Results: LPS-induced nitric oxide formation was reduced by 5AZ treatment in RAW264.7 cells. LPS-induced iNOS mRNA and protein inductions were blocked by 5AZ treatment. Next, the effect of 5AZ on IRF1, a regulator of iNOS, was examined. IRF-1 was dramatically increased by LPS to peak at 4 hr and then reduced to basal level. In the presence of MG132, a proteasome inhibitor, IRF-1 protein sustained maximal level without degradation. On the other hand, IRF-1 protein was significantly highly maintained by 5AZ treatment. In animal study, there were significant improvements in ejection fraction (53.25 ± 2.55 % vs. 62.50 ± 7.2 %, p < 0.05) and cardiac fibrosis (28.85 ± 5.44% vs. 16.57 ± 6.30%, p < 0.05). Collectively, 5AZ inhibited iNOS induction by modulation of IRF-1 in the activated macrophages to preserved cardiac function and fibrosis after MI. Conclusion: 5AZ protected post-MI injury by regulation of IRF-1 kinetics to modulate macrophages in infarcted myocardium. This study suggested 5AZ as a novel therapeutic intervention for cardiac repair.

scholarly journals Cytokine and Nitric Oxide-Dependent Gene Regulation in Islet Endocrine and Nonendocrine Cells

Function ◽  
2021 ◽  
Author(s):  
Jennifer S Stancill ◽  
Moujtaba Y Kasmani ◽  
Achia Khatun ◽  
Weiguo Cui ◽  
John A Corbett
Keyword(s):  
Nitric Oxide ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Endocrine Cells ◽  
Physiological Role ◽  
Cytokine Signaling ◽  
Inos Mrna ◽  
Β Cells ◽  
Β Cell ◽  
Shock Proteins

Abstract While exposure to inflammatory cytokines is thought to contribute to pancreatic β-cell damage during diabetes, primarily because cytokine-induced nitric oxide impairs β-cell function and causes cell death with prolonged exposure, we hypothesize that there is a physiological role for cytokine signaling that functions to protect β-cells from a number of environmental stresses. This hypothesis is derived from the knowledge that β-cells are essential for survival even though they have a limited capacity to replicate, yet they are exposed to high cytokine levels during infection as most of the pancreatic blood flow is directed to islets. Here, mouse islets were subjected to single-cell RNA sequencing following 18-hr cytokine exposure. Treatment with IL-1β and IFN-γ leads to expression of inducible nitric oxide synthase (iNOS) mRNA and antiviral and immune-associated genes as well as repression of islet identity factors in a subset of β- and non-β endocrine cells in a nitric oxide-independent manner. Nitric oxide-dependent expression of genes encoding heat shock proteins was observed in both β- and non-β endocrine cells. Interestingly, cells with high expression of heat shock proteins failed to increase antiviral and immune-associated genes, suggesting that nitric oxide may be an internal “off switch” to prevent the negative effects of prolonged cytokine signaling in islet endocrine cells. We found no evidence for pro-apoptotic gene expression following 18-hr cytokine exposure. Our findings suggest that the primary functions of cytokines and nitric oxide are to protect islet endocrine cells from damage, and only when regulation of cytokine signaling is lost does irreversible damage occur.

scholarly journals Comparing the protective effects of resveratrol, curcumin and sulforaphane against LPS/IFN-γ-mediated inflammation in doxorubicin-treated macrophages

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Haidy A. Saleh ◽  
Eman Ramdan ◽  
Mohey M. Elmazar ◽  
Hassan M. E. Azzazy ◽  
Anwar Abdelnaser
Keyword(s):  
Nitric Oxide ◽  
Inflammatory Response ◽  
Raw 264.7 Cells ◽  
Inos Mrna ◽  
No Production ◽  
Raw 264.7 ◽  
Mrna Levels ◽  
Protective Effects ◽  
Tnf Α ◽  
Ifn Γ

AbstractDoxorubicin (DOX) chemotherapy is associated with the release of inflammatory cytokines from macrophages. This has been suggested to be, in part, due to DOX-mediated leakage of endotoxins from gut microflora, which activate Toll-like receptor 4 (TLR4) signaling in macrophages, causing severe inflammation. However, the direct function of DOX on macrophages is still unknown. In the present study, we tested the hypothesis that DOX alone is incapable of stimulating inflammatory response in macrophages. Then, we compared the anti-inflammatory effects of curcumin (CUR), resveratrol (RES) and sulforaphane (SFN) against lipopolysaccharide/interferon-gamma (LPS/IFN-γ)-mediated inflammation in the absence or presence of DOX. For this purpose, RAW 264.7 cells were stimulated with LPS/IFN-γ (10 ng/mL/10 U/mL) in the absence or presence of DOX (0.1 µM). Our results showed that DOX alone is incapable of stimulating an inflammatory response in RAW 264.7 macrophages. Furthermore, after 24 h of incubation with LPS/IFN-γ, a significant increase in tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) mRNA levels was observed. Similarly, nitric oxide (NO) production and TNF-α and IL-6 protein levels were significantly upregulated. Moreover, in LPS/IFN-γ-treated macrophages, the microRNAs (miRNAs) miR-146a, miR-155, and miR-21 were significantly overexpressed. Interestingly, upon testing CUR, RES, and SFN against LPS/IFN-γ-mediated inflammation, only SFN was able to significantly reverse the LPS/IFN-γ-mediated induction of iNOS, TNF-α and IL-6 and attenuate miR-146a and miR-155 levels. In conclusion, SFN, at the transcriptional and posttranscriptional levels, exhibits potent immunomodulatory action against LPS/IFN-γ-stimulated macrophages, which may indicate SFN as a potential treatment for DOX-associated inflammation.

scholarly journals Florigen governs shoot regeneration

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yaarit Kutsher ◽  
Michal Fisler ◽  
Adi Faigenboim ◽  
Moshe Reuveni
Keyword(s):  
Nitric Oxide ◽  
Shoot Regeneration ◽  
Ectopic Expression ◽  
Flowering Plants ◽  
Regeneration Capacity ◽  
No Donor ◽  
Tobacco Leaves ◽  
Age Related ◽  
Delayed Flowering

AbstractIt is widely known that during the reproductive stage (flowering), plants do not root well. Most protocols of shoot regeneration in plants utilize juvenile tissue. Adding these two realities together encouraged us to study the role of florigen in shoot regeneration. Mature tobacco tissue that expresses the endogenous tobacco florigen mRNA regenerates poorly, while juvenile tissue that does not express the florigen regenerates shoots well. Inhibition of Nitric Oxide (NO) synthesis reduced shoot regeneration as well as promoted flowering and increased tobacco florigen level. In contrast, the addition of NO (by way of NO donor) to the tissue increased regeneration, delayed flowering, reduced tobacco florigen mRNA. Ectopic expression of florigen genes in tobacco or tomato decreased regeneration capacity significantly. Overexpression pear PcFT2 gene increased regeneration capacity. During regeneration, florigen mRNA was not changed. We conclude that florigen presence in mature tobacco leaves reduces roots and shoots regeneration and is the possible reason for the age-related decrease in regeneration capacity.

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