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Cytokine Jan 2019Interleukin (IL)-29 is known to modulate immune functions of monocytes or macrophages. In this study, we investigated the effect and its underlying mechanism of IL-29 on...
Interleukin (IL)-29 is known to modulate immune functions of monocytes or macrophages. In this study, we investigated the effect and its underlying mechanism of IL-29 on receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis using murine macrophage cell line RAW264.7 cells and bone-marrow-derived monocyte/macrophage precursor cells (BMMs), and human peripheral blood mononuclear cells (PBMCs). In response to human recombinant IL-29, cell viability and apoptosis were assessed by Cell Counting Kit-8 and flow cytometry; the osteoclast formation and activity by tartrate-resistant acid phosphatase (TRAP) staining and pit formation assay, respectively; the expression and activation of molecules that associated with osteoclastogenesis by real time-PCR, immunoblotting or immunofluorescent analysis. IL-28 receptor α (IL-28Rα), a specific receptor of IL-29 was expressed on RAW264.7 cells. Although IL-29 did not affect the viability and apoptosis of RAW264.7 cells, it inhibited multinucleated cells in the differentiation of osteoclastogenesis, the bone-resorbing activity of mature osteoclasts and osteoclastic specific genes expression including TRAP, cathepsin K (CTSK), nuclear factor of activated T-cells, cytoplasmic 1 (NFATc1), C-Fos and matrix metallopeptidase 9 (MMP-9). This inhibitory effect of IL-29 was confirmed on BMMs and PBMCs and mediated via IL-28Rα through the activation of Stat1 and 3 and the suppression of nuclear factor kappa B (NF-κB) and NFATc1 nuclear translocation in RAW264.7 cells. In conclusion, IL-29 inhibited osteoclastogenesis via activation of STAT signaling pathway, prevention of NF-κB activation and NFATc1 translocation, and suppression of downstream osteoclastogenic genes expression.
Topics: Animals; Cell Differentiation; Cell Line; Humans; Interferons; Interleukins; Leukocytes, Mononuclear; MAP Kinase Signaling System; Mice; Mice, Inbred C57BL; NF-kappa B; NFATC Transcription Factors; Osteoclasts; Osteogenesis; RANK Ligand; RAW 264.7 Cells; STAT Transcription Factors; Signal Transduction
PubMed: 30001863
DOI: 10.1016/j.cyto.2018.06.032 -
Bulletin of Experimental Biology and... Feb 2017The content of myeloid stem CFU in bone marrow karyocytes from the tibial bone of C57Bl/6 mice was evaluated after a 30-day Bion-M1 pace flight/ground control experiment...
The content of myeloid stem CFU in bone marrow karyocytes from the tibial bone of C57Bl/6 mice was evaluated after a 30-day Bion-M1 pace flight/ground control experiment and subsequent 7-day recovery period. After the space flight, we observed a significant decrease in the number of erythroid progenitors in the bone marrow, including common myeloid precursor - granulocyte, erythrocyte, monocyte/macrophage, megakaryocyte CFU. After 7-day readaptation, CFU level in flight animals did not recover completely. In the ground control, the count of erythroid burst-forming units was higher than in vivarium animals. Comparison of the changes observed in fight and ground experiments demonstrated effects associated space flight factors and manifesting in suppression of the bone marrow erythropoiesis.
Topics: Adaptation, Physiological; Animals; Bone Marrow; Cell Count; Erythrocytes; Erythropoiesis; Granulocytes; Hematopoietic Stem Cells; Male; Megakaryocytes; Mice; Mice, Inbred C57BL; Monocytes; Space Flight; Spacecraft; Tibia; Weightlessness
PubMed: 28243916
DOI: 10.1007/s10517-017-3647-8 -
Zhonghua Xue Ye Xue Za Zhi = Zhonghua... May 2023To investigate the effect of the AML1-ETO (AE) fusion gene on the biological function of U937 leukemia cells by establishing a leukemia cell model that induces AE...
To investigate the effect of the AML1-ETO (AE) fusion gene on the biological function of U937 leukemia cells by establishing a leukemia cell model that induces AE fusion gene expression. The doxycycline (Dox) -dependent expression of the AE fusion gene in the U937 cell line (U937-AE) were established using a lentivirus vector system. The Cell Counting Kit 8 methods, including the PI and sidanilide induction, were used to detect cell proliferation, cell cycle-induced differentiation assays, respectively. The effect of the AE fusion gene on the biological function of U937-AE cells was preliminarily explored using transcriptome sequencing and metabonomic sequencing. ①The Dox-dependent Tet-on regulatory system was successfully constructed to regulate the stable AE fusion gene expression in U937-AE cells. ②Cell proliferation slowed down and the cell proliferation rate with AE expression (3.47±0.07) was lower than AE non-expression (3.86 ± 0.05) after inducing the AE fusion gene expression for 24 h (<0.05). The proportion of cells in the G(0)/G(1) phase in the cell cycle increased, with AE expression [ (63.45±3.10) %) ] was higher than AE non-expression [ (41.36± 9.56) %] (<0.05). The proportion of cells expressing CD13 and CD14 decreased with the expression of AE. The AE negative group is significantly higher than the AE positive group (<0.05). ③The enrichment analysis of the transcriptome sequencing gene set revealed significantly enriched quiescence, nuclear factor kappa-light-chain-enhancer of activated B cells, interferon-α/γ, and other inflammatory response and immune regulation signals after AE expression. ④Disorder of fatty acid metabolism of U937-AE cells occurred under the influence of AE. The concentration of the medium and short-chain fatty acid acylcarnitine metabolites decreased in cells with AE expressing, propionyl L-carnitine, wherein those with AE expression (0.46±0.13) were lower than those with AE non-expression (1.00±0.27) (<0.05). The metabolite concentration of some long-chain fatty acid acylcarnitine increased in cells with AE expressing tetradecanoyl carnitine, wherein those with AE expression (1.26±0.01) were higher than those with AE non-expression (1.00±0.05) (<0.05) . This study successfully established a leukemia cell model that can induce AE expression. The AE expression blocked the cell cycle and inhibited cell differentiation. The gene sets related to the inflammatory reactions was significantly enriched in U937-AE cells that express AE, and fatty acid metabolism was disordered.
Topics: Humans; U937 Cells; RUNX1 Translocation Partner 1 Protein; Leukemia; Core Binding Factor Alpha 2 Subunit; Oncogene Proteins, Fusion; Leukemia, Myeloid, Acute
PubMed: 37550185
DOI: 10.3760/cma.j.issn.0253-2727.2023.05.003 -
Respiratory Research Mar 2013Marked accumulation of alveolar macrophages (AM) conferred by apoptosis resistance has been implicated in pathogenesis of chronic obstructive pulmonary disease (COPD)....
BACKGROUND
Marked accumulation of alveolar macrophages (AM) conferred by apoptosis resistance has been implicated in pathogenesis of chronic obstructive pulmonary disease (COPD). Apoptosis inhibitor of macrophage (AIM), has been shown to be produced by mature tissue macrophages and AIM demonstrates anti-apoptotic property against multiple apoptosis-inducing stimuli. Accordingly, we attempt to determine if AIM is expressed in AM and whether AIM is involved in the regulation of apoptosis in the setting of cigarette smoke extract (CSE) exposure.
METHODS
Immunohistochemical evaluations of AIM were performed. Immunostaining was assessed by counting total and positively staining AM numbers in each case (n = 5 in control, n = 5 in non-COPD smoker, n = 5 in COPD). AM were isolated from bronchoalveolar lavage fluid (BALF). The changes of AIM expression levels in response to CSE exposure in AM were evaluated. Knock-down of anti-apoptotic Bcl-xL was mediated by siRNA transfection. U937 monocyte-macrophage cell line was used to explore the anti-apoptotic properties of AIM.
RESULTS
The numbers of AM and AIM-positive AM were significantly increased in COPD lungs. AIM expression was demonstrated at both mRNA and protein levels in isolated AM, which was enhanced in response to CSE exposure. AIM significantly increased Bcl-xL expression levels in AM and Bcl-xL was involved in a part of anti-apoptotic mechanisms of AIM in U937 cells in the setting of CSE exposure.
CONCLUSIONS
These results suggest that AIM expression in association with cigarette smoking may be involved in accumulation of AM in COPD.
Topics: Aged; Antigens, CD; Antigens, Differentiation, T-Lymphocyte; Apoptosis Regulatory Proteins; Bronchoalveolar Lavage Fluid; Cells, Cultured; Female; Gene Expression Regulation; HEK293 Cells; Humans; Lectins, C-Type; Macrophages, Alveolar; Male; Middle Aged; Pulmonary Disease, Chronic Obstructive; U937 Cells
PubMed: 23497247
DOI: 10.1186/1465-9921-14-30 -
Journal of Experimental & Clinical... Nov 2012Epidemiological studies revealed significantly lower mortality rates in cancer patients receiving cardiac glycosides, which turned on interest in the anticancer...
BACKGROUND
Epidemiological studies revealed significantly lower mortality rates in cancer patients receiving cardiac glycosides, which turned on interest in the anticancer properties of these drugs. However, cardiac glycosides have also been shown to stimulate cell growth in several cell types. In the present investigation we analyzed the pro-death and pro-survival properties of ouabain in the human lymphoma derived cell line U937.
METHODS
ROS, intracellular Ca++, cell cycle were evaluated by loading the cells with fluorescent probes under cytofluorimetry. Cell counts and evaluation of trypan blue-excluding cells were performed under optic microscope. Protein detection was done by specific antibodies after protein separation from cellular lysates by SDS-PAGE and transfer blot.
RESULTS
High doses of ouabain cause ROS generation, elevation of [Ca++]i and death of lymphoma derived U937 cells. Lower doses of OUA activate a survival pathway in which plays a role the Na+/Ca++-exchanger (NCX), active in the Ca++ influx mode rather than in the Ca++ efflux mode. Also p38 MAPK plays a pro-survival role. However, the activation of this MAPK does not appear to depend on NCX.
CONCLUSION
This investigation shows that the cardiac glycoside OUA is cytotoxic also for the lymphoma derived cell line U937 and that can activate a survival pathway in which are involved NCX and p38 MAPK. These molecules can represent potential targets of combined therapy.
Topics: Calcium; Cell Death; Cell Survival; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Lymphoma; Ouabain; Reactive Oxygen Species; Sodium-Calcium Exchanger; U937 Cells; p38 Mitogen-Activated Protein Kinases
PubMed: 23153195
DOI: 10.1186/1756-9966-31-95 -
Experimental Parasitology Feb 2019The intraerythrocytic malaria parasite digests haemoglobin to provide amino acids for metabolism and releases toxic haem that is sequestered into haemozoin, a non-toxic,...
The intraerythrocytic malaria parasite digests haemoglobin to provide amino acids for metabolism and releases toxic haem that is sequestered into haemozoin, a non-toxic, insoluble, crystalline pigment. Following erythrocyte rupture, haemozoin is released into circulation and phagocytosed by monocytes. Phagocytosed haemozoin and antimalarial drugs have both been reported to modulate monocyte functions. This study determined the effects of therapeutic concentrations of seven antimalarial drugs; amodiaquine, artemisinin, chloroquine, doxycycline, primaquine, pyrimethamine and quinine, on the phagocytosis of β-haematin (synthetic haemozoin) by two monocytic cell lines, J774A.1 and U937, and human peripheral blood mononuclear cells. A novel spectrophotometric method based on the absorbance (O.D 400 nm) of alkali/SDS treated monocytes containing β-haematin was developed to complement counting phagocytosis with microscopy. The method has potential use for the large scale screening of monocyte phagocytic activity. Artemisinin, quinine, primaquine and pyrimethamine activated β-haematin phagocytosis by 12% or more, whereas amodiaquine, chloroquine and doxycyline inhibited β-haematin phagocytosis. In contrast, antimalarial drugs had minimal inhibitory effects on the phagocytosis of latex beads with only quinine resulting in more than 20% inhibition. Antimalarial drugs appear to alter monocyte phagocytic activity which has implications for the treatment, pathogenicity and adjunct therapies for malaria.
Topics: Amodiaquine; Animals; Antimalarials; Artemisinins; Cell Count; Cell Line; Chloroquine; Doxycycline; Electron Probe Microanalysis; Heme; Hemeproteins; Humans; Leukocytes, Mononuclear; Mice; Microscopy, Electron, Scanning; Microscopy, Electron, Transmission; Monocytes; Peroxidase; Phagocytosis; Primaquine; Pyrimethamine; Quinine; Spectrophotometry; Temperature; U937 Cells
PubMed: 30562480
DOI: 10.1016/j.exppara.2018.12.002 -
Pharmacological Research Jul 2019Poly(ADP-ribose) polymerase (PARP) is involved in the pathogenesis of cell dysfunction, inflammation and organ failure during septic shock. The goal of the current study...
The PARP inhibitor olaparib exerts beneficial effects in mice subjected to cecal ligature and puncture and in cells subjected to oxidative stress without impairing DNA integrity: A potential opportunity for repurposing a clinically used oncological drug for the experimental therapy of sepsis.
Poly(ADP-ribose) polymerase (PARP) is involved in the pathogenesis of cell dysfunction, inflammation and organ failure during septic shock. The goal of the current study was to investigate the efficacy and safety of the clinically approved PARP inhibitor olaparib in experimental models of oxidative stress in vitro and in sepsis in vivo. In mice subjected to cecal ligation and puncture (CLP) organ injury markers, circulating and splenic immune cell distributions, circulating mediators, DNA integrity and survival was measured. In U937 cells subjected to oxidative stress, cellular bioenergetics, viability and DNA integrity were measured. Olaparib was used to inhibit PARP. The results show that in adult male mice subjected to CLP, olaparib (1-10 mg/kg i.p.) improved multiorgan dysfunction. Olaparib treatment reduced the degree of bacterial CFUs. Olaparib attenuated the increases in the levels of several circulating mediators in the plasma. In the spleen, the number of CD4+ and CD8+ lymphocytes were reduced in response to CLP; this reduction was inhibited by olaparib treatment. Treg but not Th17 lymphocytes increased in response to CLP; these cell populations were reduced in sepsis when the animals received olaparib. The Th17/Treg ratio was lower in CLP-olaparib group than in the CLP control group. Analysis of miRNA expression identified a multitude of changes in spleen and circulating white blood cell miRNA levels after CLP; olaparib treatment selectively modulated these responses. Olaparib extended the survival rate of mice subjected to CLP. In contrast to males, in female mice olaparib did not have significant protective effects in CLP. In aged mice olaparib exerted beneficial effects that were less pronounced than the effects obtained in young adult males. In in vitro experiments in U937 cells subjected to oxidative stress, olaparib (1-100 μM) inhibited PARP activity, protected against the loss of cell viability, preserved NAD levels and improved cellular bioenergetics. In none of the in vivo or in vitro experiments did we observe any adverse effects of olaparib on nuclear or mitochondrial DNA integrity. In conclusion, olaparib improves organ function and extends survival in septic shock. Repurposing and eventual clinical introduction of this clinically approved PARP inhibitor may be warranted for the experimental therapy of septic shock.
Topics: Animals; Anti-Inflammatory Agents; Cecum; Cytokines; DNA; Drug Repositioning; Female; Humans; Ligation; Liver; Lung; Lymphocyte Count; Male; Mice, Inbred C57BL; Oxidative Stress; Phthalazines; Piperazines; Poly(ADP-ribose) Polymerase Inhibitors; Punctures; Sepsis; Spleen; U937 Cells
PubMed: 31071432
DOI: 10.1016/j.phrs.2019.104263 -
Journal of Endourology May 1999Escherichia coli is the bacterium most commonly isolated from the urine of patients with urinary tract infection (UTI). Recurrent episodes of UTI lead to renal... (Comparative Study)
Comparative Study
BACKGROUND
Escherichia coli is the bacterium most commonly isolated from the urine of patients with urinary tract infection (UTI). Recurrent episodes of UTI lead to renal interstitial scarring. In interstitial fibrosis and scarring, infiltration of mononuclear cells has been reported to play a key role.
MATERIALS AND METHODS
We evaluated the effect of two strains of E. coli--the pathogenic BH-5 and the plasmidless, nonfimbriated HB-101-on human monocyte and murine macrophage apoptosis.
RESULTS
E. coli BH-5 enhanced apoptosis in a time- and dose-dependent manner. It also promoted necrosis in a time- and dose-dependent manner. Strain HB-101 promoted monocyte apoptosis in a dose-dependent manner. However, the magnitude of HB-101-induced monocyte apoptosis was lower than BH-5-induced macrophage apoptosis.
CONCLUSION
The ability of E. coli to induce apoptosis may contribute to its virulence and play a role in renal interstitial scarring.
Topics: Animals; Apoptosis; Cell Count; Cicatrix; DNA; DNA Fragmentation; Electrophoresis, Agar Gel; Escherichia coli; Escherichia coli Infections; Fibrosis; Humans; Macrophages, Peritoneal; Mice; Monocytes; Necrosis; U937 Cells; Urinary Tract Infections; Virulence
PubMed: 10405905
DOI: 10.1089/end.1999.13.273 -
Journal of Ethnopharmacology Jan 2009Korean red ginseng (KRG, Panax ginseng C.A. Meyer Radix rubra) has been used to treat various diseases including cancer. However, the molecular mechanisms responsible...
AIM OF THIS STUDY
Korean red ginseng (KRG, Panax ginseng C.A. Meyer Radix rubra) has been used to treat various diseases including cancer. However, the molecular mechanisms responsible for KRG extract induced apoptosis and telomerase inhibition remain unclear.
MATERIALS AND METHODS
The hot water extract from KRG was used to evaluate the mechanism of induction of apoptosis in U937 human leukemia cells and its effects on cyclooxgenase-2 (COX-2) and telomerase activity.
RESULTS
KRG extract treatment to U937 cells resulted in growth inhibition and induction of apoptosis in a concentration-dependent manner as measured by hemacytometer counts, MTT assay, fluorescence microscopy, agarose gel electrophoresis and flow cytometry analysis. The increase in apoptosis was associated with the down-regulation of antiapoptotic Bcl-2, Bcl-X(L), and IAPs family members, and the activation of caspase-3. KRG extract treatment also decreased the expression levels of COX-2 and inducible nitric oxide synthase. Furthermore, KRG extract treatment progressively down-regulated the expression of human telomerase reverse transcriptase, a main determinant of the telomerase enzymatic activity, with inhibiting the expression of c-Myc in a concentration-dependent manner.
CONCLUSIONS
These results provide important new insights into the possible molecular mechanisms of the anticancer activity of KRG extract.
Topics: Antineoplastic Agents, Phytogenic; Apoptosis; Cyclooxygenase 2; Dose-Response Relationship, Drug; Electrophoresis, Agar Gel; Flow Cytometry; Gene Expression Regulation, Neoplastic; Humans; Lymphoma, Large B-Cell, Diffuse; Medicine, Korean Traditional; Microscopy, Fluorescence; Panax; Plant Extracts; Proto-Oncogene Proteins c-myc; Telomerase; U937 Cells
PubMed: 19041934
DOI: 10.1016/j.jep.2008.10.038 -
Arteriosclerosis, Thrombosis, and... Jun 2005Apoptotic cell death has been demonstrated in advanced human atherosclerotic plaques. Apoptotic cells (ACs) should be rapidly removed by macrophages, otherwise secondary...
OBJECTIVE
Apoptotic cell death has been demonstrated in advanced human atherosclerotic plaques. Apoptotic cells (ACs) should be rapidly removed by macrophages, otherwise secondary necrosis occurs, which in turn elicits inflammatory responses and plaque progression. Therefore, we investigated the efficiency of phagocytosis of ACs by macrophages in atherosclerosis.
METHODS AND RESULTS
Human endarterectomy specimens and human tonsils were costained for CD68 (macrophages) and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) (apoptosis). Free and phagocytized ACs were counted in both tissues. The ratio of free versus phagocytized AC was 19-times higher in human atherosclerotic plaques as compared with human tonsils, indicating a severe defect in clearance of AC. Impaired phagocytosis of AC was also detected in plaques from cholesterol-fed rabbits and did not further change with plaque progression. In vitro experiments with J774 or peritoneal mouse macrophages showed that several factors caused impaired phagocytosis of AC including cytoplasmic overload of macrophages with indigestible material (beads), free radical attack, and competitive inhibition among oxidized red blood cells, oxidized low-density lipoprotein and ACs for the same receptor(s) on the macrophage.
CONCLUSIONS
Our data demonstrate that phagocytosis of ACs is impaired in atherosclerotic plaques, which is at least partly attributed to oxidative stress and cytoplasmic saturation with indigestible material.
Topics: Aged; Animals; Apoptosis; Carotid Artery Diseases; Cytoplasm; Erythrocytes; Female; Humans; Lipoproteins, LDL; Macrophages, Peritoneal; Male; Mice; Necrosis; Oxidative Stress; Palatine Tonsil; Phagocytosis; Rabbits; U937 Cells
PubMed: 15831805
DOI: 10.1161/01.ATV.0000166517.18801.a7