scholarly journals IL-10 and class 1 histone deacetylases act synergistically and independently on the secretion of proinflammatory mediators in alveolar macrophages

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0245169
Author(s):  
Brent A. Stanfield ◽  
Todd Purves ◽  
Scott Palmer ◽  
Bruce Sullenger ◽  
Karen Welty-Wolf ◽  
...  
Keyword(s):  
Alveolar Macrophages ◽  
Histone Deacetylases ◽  
Pulmonary Inflammation ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Synthesis Inhibitor ◽  
Proinflammatory Mediators ◽  
Chemokine Signaling ◽  
Class 1 ◽  
Anti Inflammatory

Introduction Anti-inflammatory cytokine IL-10 suppresses pro-inflammatory IL-12b expression after Lipopolysaccharide (LPS) stimulation in colonic macrophages, as part of the innate immunity Toll-Like Receptor (TLR)-NF-κB activation system. This homeostatic mechanism limits excess inflammation in the intestinal mucosa, as it constantly interacts with the gut flora. This effect is reversed with Histone Deacetylase 3 (HDAC3), a class I HDAC, siRNA, suggesting it is mediated through HDAC3. Given alveolar macrophages’ prominent role in Acute Lung Injury (ALI), we aim to determine whether a similar regulatory mechanism exists in the typically sterile pulmonary microenvironment. Methods Levels of mRNA and protein for IL-10, and IL-12b were determined by qPCR and ELISA/Western Blot respectively in naïve and LPS-stimulated alveolar macrophages. Expression of the NF-κB intermediaries was also similarly assessed. Experiments were repeated with AS101 (an IL-10 protein synthesis inhibitor), MS-275 (a selective class 1 HDAC inhibitor), or both. Results LPS stimulation upregulated all proinflammatory mediators assayed in this study. In the presence of LPS, inhibition of IL-10 and/or class 1 HDACs resulted in both synergistic and independent effects on these signaling molecules. Quantitative reverse-transcriptase PCR on key components of the TLR4 signaling cascade demonstrated significant diversity in IL-10 and related gene expression in the presence of LPS. Inhibition of IL-10 secretion and/or class 1 HDACs in the presence of LPS independently affected the transcription of MyD88, IRAK1, Rela and the NF-κB p50 subunit. Interestingly, by quantitative ELISA inhibition of IL-10 secretion and/or class 1 HDACs in the presence of LPS independently affected the secretion of not only IL-10, IL-12b, and TNFα, but also proinflammatory mediators CXCL2, IL-6, and MIF. These results suggest that IL-10 and class 1 HDAC activity regulate both independent and synergistic mechanisms of proinflammatory cytokine/chemokine signaling. Conclusions Alveolar macrophages after inflammatory stimulation upregulate both IL-10 and IL-12b production, in a highly class 1 HDAC-dependent manner. Class 1 HDACs appear to help maintain the balance between the pro- and anti-inflammatory IL-12b and IL-10 respectively. Class 1 HDACs may be considered as targets for the macrophage-initiated pulmonary inflammation in ALI in a preclinical setting.

scholarly journals Social Fear Memory Requires Two Stages of Protein Synthesis in Mice

2020 ◽  
Vol 21 (15) ◽  
pp. 5537
Author(s):  
Johannes Kornhuber ◽  
Iulia Zoicas
Keyword(s):  
Protein Synthesis ◽  
Temporal Dynamics ◽  
De Novo ◽  
Fear Memory ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Synthesis Inhibitor ◽  
Systemic Injection ◽  
Social Fear ◽  
Two Stages

It is well known that long-term consolidation of newly acquired information, including information related to social fear, require de novo protein synthesis. However, the temporal dynamics of protein synthesis during the consolidation of social fear memories is unclear. To address this question, mice received a single systemic injection with the protein synthesis inhibitor, anisomycin, at different time-points before or after social fear conditioning (SFC), and memory was assessed 24 h later. We showed that anisomycin impaired the consolidation of social fear memories in a time-point-dependent manner. Mice that received anisomycin 20 min before, immediately after, 6 h, or 8 h after SFC showed reduced expression of social fear, indicating impaired social fear memory, whereas anisomycin caused no effects when administered 4 h after SFC. These results suggest that consolidation of social fear memories requires two stages of protein synthesis: (1) an initial stage starting during or immediately after SFC, and (2) a second stage starting around 6 h after SFC and lasting for at least 5 h.

scholarly journals Antinociceptive and Anti-Inflammatory Effects of Total Alkaloid Extract fromFumaria capreolata

2015 ◽  
Vol 2015 ◽  
pp. 1-7 ◽  
Author(s):  
Noureddine Bribi ◽  
Francesca Algieri ◽  
Alba Rodriguez-Nogales ◽  
Jose Garrido-Mesa ◽  
Teresa Vezza ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Inflammatory Diseases ◽  
Potential Candidate ◽  
Dependent Manner ◽  
Proinflammatory Mediators ◽  
Total Alkaloids ◽  
Aerial Parts ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Fumaria Capreolata

Fumaria capreolatais used in traditional medicine in North Africa for its gastrointestinal and anti-inflammatory activities. The present study investigates the effects of total alkaloids extracted from the aerial parts ofFumaria capreolata(AFC) on LPS-induced production of proinflammatory mediators (IL-6, IL-1β, iNOS, TNF-α, COX-2, and MIP-2) in RAW264.7 cells. AFC significantly reduced the inflammatory response inhibiting the production of nitric oxide (NO) and IL-6 in a dose-dependent manner, without affecting the viability of cells, and downregulated mRNA expression of proinflammatory key players: IL-6, IL-1β, iNOS, TNF-α, and COX-2. AFC antinociceptive and anti-inflammatory properties were also evaluated on the acetic acid- and formalin-induced pain models in mice. AFC oral administration significantly inhibited acetic acid-induced writhes and reduced formalin-induced paw licking time. Therefore, AFC may be a potential candidate for the treatment of inflammatory diseases, such as colitis and arthritis.

scholarly journals Signal transduction for chemotaxis and haptotaxis by matrix molecules in tumor cells.

1990 ◽  
Vol 110 (4) ◽  
pp. 1427-1438 ◽  
Author(s):  
S Aznavoorian ◽  
M L Stracke ◽  
H Krutzsch ◽  
E Schiffmann ◽  
L A Liotta
Keyword(s):  
Protein Synthesis ◽  
Matrix Protein ◽  
Type Iv Collagen ◽  
De Novo ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Recognition Sequence ◽  
Synthesis Inhibitor ◽  
Concentration Dependent Manner ◽  
Type Iv

Transduction of signals initiating motility by extracellular matrix (ECM) molecules differed depending on the type of matrix molecule and whether the ligand was in solution or bound to a substratum. Laminin, fibronectin, and type IV collagen stimulated both chemotaxis and haptotaxis of the A2058 human melanoma cell line. Peak chemotactic responses were reached at 50-200 nM for laminin, 50-100 nM for fibronectin, and 200-370 nM for type IV collagen. Checkerboard analysis of each attractant in solution demonstrated a predominantly directional (chemotactic) response, with a minor chemokinetic component. The cells also migrated in a concentration-dependent manner to insoluble step gradients of substratum-bound attractant (haptotaxis). The haptotactic responses reached maximal levels at coating concentrations of 20 nM for laminin and type IV collagen, and from 30 to 45 nM for fibronectin. Pretreatment of cells with the protein synthesis inhibitor, cycloheximide (5 micrograms/ml), resulted in a 5-30% inhibition of both chemotactic and haptotactic responses to each matrix protein, indicating that de novo protein synthesis was not required for a significant motility response. Pretreatment of cells with 50-500 micrograms/ml of synthetic peptides containing the fibronectin cell-recognition sequence GRGDS resulted in a concentration-dependent inhibition of fibronectin-mediated chemotaxis and haptotaxis (70-80% inhibition compared to control motility); negative control peptide GRGES had only a minimal effect. Neither GRGDS nor GRGES significantly inhibited motility to laminin or type IV collagen. Therefore, these results support a role for the RGD-directed integrin receptor in both types of motility response to fibronectin. After pretreatment with pertussis toxin (PT), chemotactic responses to laminin, fibronectin, and type IV collagen were distinctly different. Chemotaxis to laminin was intermediate in sensitivity; chemotaxis to fibronectin was completely insensitive; and chemotaxis to type IV collagen was profoundly inhibited by PT. In marked contrast to the inhibition of chemotaxis, the hepatotactic responses to all three ligands were unaffected by any of the tested concentrations of PT. High concentrations of cholera toxin (CT; 10 micrograms/ml) or the cAMP analogue, 8-Br-cAMP (0.5 mM), did not significantly affect chemotactic or haptotactic motility to any of the attractant proteins, ruling out the involvement of cAMP in the biochemical pathway initiating motility in these cells. The sensitivity of chemotaxis induced by laminin and type IV collagen, but not fibronectin, to PT indicates the involvement of a PT-sensitive G protein in transduction of the signals initiating motility to soluble laminin and type IV collagen.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Duchesnea indica Extract Attenuates Coal Fly Ash-Induced Inflammation in Murine Alveolar Macrophages through the NF-Kappa B Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
H. M. Arif Ullah ◽  
Yuan Yee Lee ◽  
Sung Dae Kim ◽  
Man Hee Rhee
Keyword(s):  
Nitric Oxide ◽  
Fly Ash ◽  
Western Blotting ◽  
Alveolar Macrophages ◽  
Coal Fly Ash ◽  
Gas Chromatography Mass Spectrometry ◽  
No Production ◽  
Proinflammatory Mediators ◽  
Duchesnea Indica ◽  
Anti Inflammatory

Duchesnea indica is known as false strawberry, is found in East Asia, and has numerous biological properties. The aim of this study was to investigate the anti-inflammatory effect of Duchesnea indica extract (DIE) on coal fly ash- (CFA-) induced inflammation in murine alveolar macrophages (MH-S). Following the induction of inflammation in MH-S cells by CFA, nitric oxide (NO) was measured to evaluate the anti-inflammatory property of DIE. Cell viability and inflammatory gene expression were analyzed using polymerase chain reaction (PCR). The inflammatory pathway in MH-S cells was determined via western blotting and immunofluorescence (IF) analysis. Finally, the major components of the DIE were identified and separated through ultra-performance liquid chromatography (UPLC) and gas chromatography-mass spectrometry (GC-MS) analysis. Our results showed that the DIE dose-dependently inhibited the CFA-induced NO production in MH-S cells. Moreover, the DIE could suppress the CFA-induced proinflammatory mediators, such as cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). In addition, the inhibitory effect of the DIE on proinflammatory cytokines, including interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α), was detected with PCR. Moreover, the effect of the DIE on the nuclear factor-κB (NF-κB) pathway in CFA-activated MH-S cells was measured via western blotting. Furthermore, the inhibition of the phosphorylated NF-κB (p-NF-κB) translocation was analyzed using IF assay. The findings of this study indicated that the DIE potentially inhibited the CFA-induced inflammation by blocking the NF-κB inflammatory signaling pathway in MH-S cells and that the DIE might contain favorable anti-inflammatory compounds which may be effective in attenuating lung inflammation.

scholarly journals Platelets instruct T reg cells and macrophages in the resolution of lung inflammation

2021 ◽  
Vol 218 (7) ◽  
Author(s):  
Rick Kapur ◽  
John W. Semple
Keyword(s):  
Alveolar Macrophages ◽  
Lung Inflammation ◽  
Potential Role ◽  
Pulmonary Inflammation ◽  
Immune Functions ◽  
Inflammatory Phenotype ◽  
Anti Inflammatory

Platelets convey important nonhemostatic immune functions; however, their potential role in resolving pulmonary inflammation remains to be determined. In this issue of JEM, Rossaint et al. (2021. J. Exp. Med. https://doi.org/10.1084/jem.20201353) reveal that platelets contribute to the resolution of pulmonary inflammation by directly recruiting T regulatory (T reg) cells to the lungs and by transcriptionally reprogramming alveolar macrophages toward an anti-inflammatory phenotype.

scholarly journals Topical Anti-Inflammatory Activity of Essential Oils of Alpinia calcarata Rosc., Its Main Constituents, and Possible Mechanism of Action

2020 ◽  
Vol 2020 ◽  
pp. 1-19
Author(s):  
Madhuvanthi Chandrakanthan ◽  
Shiroma M. Handunnetti ◽  
Galbada Sirimal Arachchige Premakumara ◽  
Selvaluxmy Kathirgamanathar
Keyword(s):  
Skin Inflammation ◽  
Inflammatory Activity ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Proinflammatory Mediators ◽  
Mouse Ear ◽  
Anti Inflammatory ◽  
Dose Dependent ◽  
Topical Analgesic

This study aimed at investigating the anti-inflammatory potential of essential oil from rhizome and leaf of Alpinia calcarata Rosc. (ACEO) with the focus of its topical anti-inflammatory activity along with its dominant compounds 1,8-cineole and α-terpineol using mouse ear edema model. ACEOs were analyzed by GC-MS. The anti-inflammatory activity was determined by studying the inhibition of overproduction of proinflammatory mediators—nitric oxide, reactive oxygen species, prostaglandins, cyclooxygenases, and cytokines induced by lipopolysaccharides in murine macrophages. Topical anti-inflammatory and antinociceptive activity was studied by 12-O-tetradecanoylphorbol-13-acetate (TPA) induced skin inflammation and formalin-induced pain model in mice, respectively. Rhizome oil has 1,8-cineole (31.08%), α-terpineol (10.31%), and fenchyl acetate (10.73%) as major compounds whereas the ACEO from leaves has 1,8-cineole (38.45%), a-terpineol (11.62%), and camphor (10%). ACEOs reduced the production of inflammatory mediators in vitro in a concentration-dependent manner. Further, ACEO and its major compounds reduced ear thickness, weight, myeloperoxidase, and cytokines significantly (p<0.01) in mouse ear. Dose-dependent reduction in flinching and licking in both the phases of pain sensation concludes the topical analgesic effect. Our findings suggest the potency of topical use of ACEOs for inflammatory disease conditions.

scholarly journals Granulocyte/macrophage colony-stimulating factor stimulates the expression of the 5-lipoxygenase-activating protein (FLAP) in human neutrophils.

1994 ◽  
Vol 179 (4) ◽  
pp. 1225-1232 ◽  
Author(s):  
M Pouliot ◽  
P P McDonald ◽  
P Borgeat ◽  
S R McColl
Keyword(s):  
De Novo ◽  
Human Neutrophils ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Colony Stimulating Factor ◽  
Synthesis Inhibitor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

The synthesis of leukotrienes in human blood neutrophils chiefly relies on the activity of two enzymes, phospholipase A2 and 5-lipoxygenase (5-LO). In turn, the activation of the 5-LO requires the participation of a recently characterized membrane-bound protein, the 5-LO-activating protein (FLAP). In this study, we have investigated conditions under which FLAP expression in neutrophils may be modulated. Of several cytokines tested, only granulocyte/macrophage colony-stimulating factor (GM-CSF) (and to a lesser extent tumor necrosis factor alpha) significantly increased expression of FLAP. GM-CSF increased FLAP mRNA steady-state levels in a time- and dose-dependent manner. The stimulatory effect of GM-CSF on FLAP mRNA was inhibited by prior treatment of the cells with the transcription inhibitor, actinomycin D, and pretreatment of the cells with the protein synthesis inhibitor, cycloheximide, failed to prevent the increase in FLAP mRNA induced by GM-CSF. The accumulation of newly synthesized FLAP, as determined by immunoprecipitation after incorporation of 35S-labeled amino acids, was also increased after incubation of neutrophils with GM-CSF. In addition, the total level of FLAP protein was increased in GM-CSF-treated neutrophils, as determined by two-dimensional gel electrophoresis, followed by Western blot. GM-CSF did not alter the stability of the FLAP protein, indicating that the effect of GM-CSF on FLAP accumulation was the consequence of increased de novo synthesis as opposed to decreased degradation of FLAP. Finally, incubation of neutrophils with the synthetic glucocorticoid dexamethasone directly stimulated the upregulation of FLAP mRNA and protein, and enhanced the effect of GM-CSF. Taken together, these data demonstrate that FLAP expression may be upmodulated after appropriate stimulation of neutrophils. The increase in FLAP expression induced by GM-CSF in inflammatory conditions could confer upon neutrophils a prolonged capacity to synthesize leukotrienes.

scholarly journals Circadian rhythm from the eye of Aplysia: temperature compensation of the effects of protein synthesis inhibitors

1980 ◽  
Vol 84 (1) ◽  
pp. 1-15
Author(s):  
J. W. Jacklet
Keyword(s):  
Protein Synthesis ◽  
Circadian Rhythm ◽  
Circadian Clock ◽  
Culture Medium ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Protein Synthesis Inhibitors ◽  
Response Curves ◽  
Synthesis Inhibitor ◽  
Maximum Phase

1. The circadian rhythm of compound action potentials (CAP) frequency recorded from the isolated eye of Aplysia in culture medium and darkness was subjected to step and pulse treatments with anisomycin, a protein synthesis inhibitor. 2. The step application of anisomycin and its continued presence in the culture medium lengthened the period of the rhythm in a dose-dependent manner. At 10(−8) M the period was increased from the normal 26.5 h to about 28 h and at 10(−7) M the period was lengthened to 31 h or longer. At 10(−6) M the rhythm was suppressed but the CAP activity continued without the cyclic variations in CAP frequency. 3. Six-hour pulses of anisomycin at 10(−6) M caused phase-dependent phase-shifts of the rhythm. Maximum phase delays of 15 h were obtained at CT (circadian time) 2 and maximum phase advances of 4 h were obtained at CT 6. The phase response curves at 12, 15 and 17 degrees C were essentially identical. 4. Anisomycin appears to act rather selectively on the circadian clock mechanism. It does not alter the CAP amplitude and duration and it does not alter the bursting pacemaker mechanism of the optic nerve CAP or central neurones. 5. The results support the hypothesis that the synthesis of a protein or polypeptide on eucaryotic ribosomes is an essential part of the circadian clock timing mechanism. The sensitivity of the clock to anisomycin is the same at three different temperatures (12, 15 and 17 degrees C) within the physiological range of temperatures for Aplysia, as expected for a clock whose period length is temperature compensated (Q10 1.02) over that same range. 6. At the critical phases of CT 1-4, anisomycin pulses often caused unusual perturbations of the rhythm. These effects are consistent with the hypothesis that the circadian rhythm is a multioscillator system.

Synchronization of porcine oocyte meiosis using cycloheximide and its application to the study of regulation by cumulus cells

2002 ◽  
Vol 14 (7) ◽  
pp. 433 ◽  
Author(s):  
J. Ye ◽  
A. P. F. Flint ◽  
K. H. S. Campbell ◽  
M. R. Luck
Keyword(s):  
Germinal Vesicle ◽  
Cumulus Cells ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Synthesis Inhibitor ◽  
Germinal Vesicle Breakdown ◽  
Nuclear Maturation ◽  
Meiotic Progression ◽  
Conventional Culture ◽  
Oocyte Meiosis

This paper describes the use of the protein synthesis inhibitor cycloheximide (CHX) to synchronize nuclear progression during meiotic maturation in porcine oocytes, and also the time-dependence of nuclear maturation on exposure of the oocyte to cumulus cells. Prior to culture, the majority of oocytes were at the germinal vesicle (GV) stage (95–100%), but distributed from GVI to GVIV (GVI 56.1 ± 9.1%, GVII 15.3 ± 1.4%, GVIII 21.5 ± 7.1%, GVIV 7.1 ± 3.5%). During culture of cumulus-enclosed oocytes (COCs) from 12 h to 48 h in a conventional culture system, all meiotic stages were represented at any time point examined, with 63.6 ± 4.2% of oocytes maturing to metaphase II (MII). Cycloheximide blocked the progression of nuclear development in a dose-dependent manner. Treatment for 12 h with CHX at 1–25 μg mL–1 resulted in 95–100% oocytes being arrested and synchronized at GVII. With >5 μg mL–1 CHX, all oocytes were arrested before germinal vesicle breakdown (GVBD) (mostly at GVIII) by 24 h. A 12 h preincubation with 5 μg mL–1 CHX followed by 24 h of further culture without CHX resulted in >80% of oocytes maturing to MII. The profile of nuclear progression during maturation revealed discrete peaks of occurrence of different meiotic stages, with GVBD at 6–12 h, metaphase I (MI) at 10–18�h and anaphase I/telophase I at 16–20 h. After 12 h preincubation with 5 μg mL–1 CHX, denuded oocytes (DOs) matured to MI as COCs. However, DOs matured to MII as normal when denuded at MI. In conclusion, CHX not only efficiently blocks and synchronizes the meiotic progression of porcine oocytes at a specific GV stage, but it also effectively synchronizes subsequent meiotic progression to MII, resulting in discrete peaks of occurrence of different meiotic stages. Using this technique, the study showed that cumulus cells are essential for oocytes to mature from MI to MII but exposure to cumulus cells must occur before MI.

scholarly journals Effect of Phosphatase Activity of the Control of Virulence Sensor (CovS) on Clindamycin-Mediated Streptolysin O Production in Group A Streptococcus

2019 ◽  
Vol 87 (12) ◽  
Author(s):  
Chuan Chiang-Ni ◽  
Huei-Chuan Tseng ◽  
Yong-An Shi ◽  
Cheng-Hsun Chiu
Keyword(s):  
Phosphatase Activity ◽  
Group A Streptococcus ◽  
Response Regulator ◽  
Protein Synthesis Inhibitor ◽  
Dependent Manner ◽  
Tissue Destruction ◽  
Synthesis Inhibitor ◽  
Content Type ◽  
Streptolysin O ◽  
Group A

ABSTRACT Severe manifestations of group A Streptococcus (GAS) infections are associated with massive tissue destruction and high mortality. Clindamycin (CLI), a bacterial protein synthesis inhibitor, is recommended for treating patients with severe invasive GAS infection. Nonetheless, the subinhibitory concentration of CLI induces the production of GAS virulent exoproteins, such as streptolysin O (SLO) and NADase, which would enhance bacterial virulence and invasiveness. A better understanding of the molecular mechanism of how CLI triggers GAS virulence factor expression will be critical to develop appropriate therapeutic approaches. The present study shows that CLI activates SLO and NADase expressions in the emm1-type CLI-susceptible wild-type strain but not in covS or control of virulence sensor (CovS) phosphatase-inactivated mutants. Supplementation with Mg2+, which is a CovS phosphatase inhibitor, inhibits the CLI-mediated SLO upregulation in a dose-dependent manner in CLI-susceptible and CLI-resistant strains. These results not only reveal that the phosphorylation of response regulator CovR is essential for responding to CLI stimuli, but also suggest that inhibiting the phosphatase activity of CovS could be a potential strategy for the treatment of invasive GAS infection with CLI.

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