The Mycoplasma pneumoniae HapE alters the cytokine profile and growth of human bronchial epithelial cells

Abstract Mycoplasma pneumoniae is one of the most common pathogenic causes of community-acquired pneumonia. Hydrogen sulfide, alanine, and pyruvate producing enzyme (HapE) is a recently discovered M. pneumoniae virulence factor that can produce H2S to promote erythrocyte lysis. However, other cytotoxic effects of HapE have not been explored. The present study examined the effects of this enzyme on normal human bronchial epithelial (NHBE) cells, in an attempt to identify additional mechanisms of M. pneumoniae pathogenesis. Recombinant HapE was purified for use in downstream assays. MTT and colony formation assays were conducted to determine the effects of HapE on cell viability and growth, while flow cytometry was used to examine changes in cell proliferation and cell cycle function. ELISA was performed to examine changes in the cytokine profile of HapE-treated cells. HapE treatment arrested NHBE cells in S phase and inhibited cell proliferation in a concentration-dependent manner. The anti-inflammatory factors interleukin (IL)-4 and IL-6 were significantly enhanced following HapE treatment. Increased secretion of pro-inflammatory factors was not observed. The effects of HapE on the respiratory epithelium may have an impact on the efficiency of host immune surveillance and pathogen elimination, and contribute to the pathogenesis of M. pneumoniae.

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Allyl Isothiocyanate Increases MRP1 Function and Expression in a Human Bronchial Epithelial Cell Line

Oxidative Medicine and Cellular Longevity ◽

10.1155/2014/547379 ◽

2014 ◽

Vol 2014 ◽

pp. 1-7 ◽

Cited By ~ 7

Author(s):

Dian-lei Wang ◽

Chen-yin Wang ◽

Yin Cao ◽

Xian Zhang ◽

Xiu-hua Tao ◽

...

Keyword(s):

Epithelial Cell ◽

Protein Expression ◽

Cell Line ◽

Allyl Isothiocyanate ◽

Bronchial Epithelial Cell ◽

Epithelial Cell Line ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Bronchial Epithelial ◽

Mrp1 Protein

Multidrug resistance-associated protein 1 (MRP1), a member of the ATP-binding cassette (ABC) superfamily of transporters, plays an important role in normal lung physiology by protecting cells against oxidative stress and toxic xenobiotics. The present study investigates the effects of allyl isothiocyanate (AITC) onMRP1mRNA and MRP1 protein expression and transporter activity in the immortalised human bronchial epithelial cell line 16HBE14o-.MRP1mRNA and MRP1 protein expression in 16HBE14o- cells that were treated with allyl isothiocyanate were analysed by real-time PCR assay and Western blotting. The transport of carboxyfluorescein, a known MRP1 substrate, was measured by functional flow cytometry to evaluate MRP1 activity. Treatment with AITC at concentrations of 5–40 μM increased MRP1 protein levels in a concentration-dependent manner. AITC treatments at concentrations of 1–40 μM caused concentration-dependent increases inMRP1mRNA levels that were up to seven times greater than the levels found in control cells. Finally, AITC treatment at concentrations of 5–40 μM significantly increased MRP1-dependent efflux in 16HBE14o- cells. These results suggest that AITC can increase the expression and activity of MRP1 in 16HBE14o- cells in a concentration-dependent manner. The upregulation of MRP1 activity and expression by AITC could produce therapeutic effects in the treatment of lung disease.

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Effects of Irbesartan on Phenotypic Alteration of Monocyte and Inflammatory Status of Hypertensive Patients with Left Ventricular Hypertrophy

10.21203/rs.3.rs-193827/v1 ◽

2021 ◽

Author(s):

Jingsi Zhang ◽

Lina Yang ◽

Yanchun Ding

Keyword(s):

Ventricular Hypertrophy ◽

Target Organ Damage ◽

Organ Damage ◽

Left Ventricular ◽

Inflammatory Factors ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Hypertensive Patients ◽

Inflammatory Status ◽

Tnf Α

Abstract Background: Circulating monocytes and tissue macrophages play complex roles in pathogenesis of hypertension and its target organ damage. In this study we observed the alterations of monocyte phenotype and inflammatory state of hypertensive patients with left ventricular hypertrophy (LVH) and the effects of Irbesartan on it. We explored new mechanisms of Irbesartan to reverse LVH and provided new targets for the prevention and treatment of hypertensive target organ damage.Methods: The CD163 and CD206 expressions on monocytes were detected as well as the IL-10 and TNF-α levels in serum of hypertensive patients with or without LVH and healthy volunteers. Furthermore, we treated monocytes of LVH group with different concentrations of Irbesartan, and then CD163, CD206, IL-10 and TNF-α were detected. Results: We found for the first time that the expression of CD163, CD206 and IL-10 of LVH group was lower than that of non-LVH group and healthy control, but the TNF-α level of LVH group was significantly higher. Irbesartan could upregulate the expression of CD163 and CD206 of hypertensive patients with LVH in a concentration-dependent manner. Irbesartan could also increase the expression of IL-10 and inhibit the expression of TNF-α in monocytes culture supernatant in a concentration-dependent manner. Conclusions: Our data suggests that inflammation was activated in hypertensive patients with LVH, the monocyte phenotype was mainly pro-inflammatory. The expression of pro-inflammatory factors increased while the expression of anti-inflammatory factors decreased. Irbesartan could alter monocyte phenotype and inflammatory status in hypertensive patients with LVH, which may be a new mechanism of Irbesartan to reverse LVH.Trail registration: The study protocols were approved by the Ethical Committee of the Second Affiliated Hospital of Dalian Medical University. Each patient signed the informed consent.

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Metabolite Profile and Antiproliferative Effects in HaCaT Cells of a Salix reticulata Extract

Planta Medica ◽

10.1055/s-0043-109098 ◽

2017 ◽

Vol 83 (14/15) ◽

pp. 1149-1158 ◽

Cited By ~ 2

Author(s):

Elisabetta Corradi ◽

Nadine Schmidt ◽

Nathalie Räber ◽

Maria De Mieri ◽

Matthias Hamburger ◽

...

Keyword(s):

Cell Proliferation ◽

Cell Migration ◽

Time Lapse ◽

Metabolite Profile ◽

Brdu Incorporation ◽

Dependent Manner ◽

Hacat Cells ◽

Meoh Extract ◽

Concentration Dependent Manner ◽

Phenolic Constituents

AbstractPhenolic constituents of Salix reticulata (Salicaceae) and antiproliferative activity of an extract and individual compounds were investigated in immortalized human non-tumorigenic keratinocytes (HaCaT). A MeOH extract from aerial parts afforded several flavonoids, including luteolin and apigenin glycosides (2–5 and 9) and catechin (1), two procyanidin fractions, and the phenolic glucosides picein (6), triandrin (7), and salicortin (8). In an adenosine triphosphate assay, the MeOH extract reduced cell viability by approximately 60 % at a concentration of 100 µg/mL. Cell proliferation was assessed with a BrdU incorporation ELISA assay. The extract inhibited proliferation of HaCaT cells in a concentration-dependent manner, with approximately 50 % inhibition at 100 µg/mL. In time-lapse assays, the extract showed distinct inhibitory effects on cell migration at concentrations of 12.5, 25, and 50 µg/mL. The activity of selected constituents was also determined. Luteolin-7-O-β-glucuronide (3) significantly inhibited cell proliferation at concentrations of 10 and 50 µM. In contrast, luteolin-7-O-β-glucopyranoside (2) and a procyanidin fraction (P1) had only weak effects, while picein (6) and salicortin (8) did not affect cell proliferation. Luteolin-7-O-β-glucuronide (10 µM) and, to a lesser extent, the procyanidin fraction (10 µg/mL) also inhibited cell migration.

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The mononucleotide-dependent, nonantisense mechanism of action of phosphodiester and phosphorothioate oligonucleotides depends upon the activity of an ecto-5′-nucleotidase

Blood ◽

10.1182/blood.v98.4.995 ◽

2001 ◽

Vol 98 (4) ◽

pp. 995-1002 ◽

Cited By ~ 23

Author(s):

Maria Koziolkiewicz ◽

Edyta Gendaszewska ◽

Maria Maszewska ◽

C. A. Stein ◽

Wojciech J. Stec

Keyword(s):

Cell Proliferation ◽

Endothelial Cells ◽

Enzymatic Degradation ◽

Umbilical Vein ◽

Cell Types ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Cytotoxic Effects ◽

Specific Effects ◽

Umbilical Vein Endothelial Cells

Many reports indicate different nonantisense yet sequence-specific effects of antisense phosphorothioate oligonucleotides. Products of enzymatic degradation of the oligonucleotides can also influence cell proliferation. The cytotoxic effects of deoxyribonucleoside-5′-phosphates (dNMPs) and their 5′-phosphorothioate analogs, deoxyribonucleoside-5′-monophosphorothioates (dNMPSs) on 4 human cell types (HeLa, HL-60, K-562, and endothelial cells) were examined, and the effects were correlated with the catabolism of these compounds. The results indicate that differences in cytotoxicity of dNMPs or dNMPSs in these cells depend upon different activity of an ecto-5′-nucleotidase. It has also been found that dNMPSs stimulate proliferation of human umbilical vein endothelial cells and HL-60 cells in a concentration-dependent manner. This stimulation might be caused by the binding of deoxynucleoside-5′-phosphorothioates to as-yet unidentified nucleotide receptor(s) at the cell surface.

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Role of Mycoplasma pneumoniae glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in mediating interactions with the human extracellular matrix

Microbiology ◽

10.1099/mic.0.048298-0 ◽

2011 ◽

Vol 157 (8) ◽

pp. 2328-2338 ◽

Cited By ~ 30

Author(s):

Roger Dumke ◽

Marius Hausner ◽

Enno Jacobs

Keyword(s):

Extracellular Matrix ◽

Mycoplasma Pneumoniae ◽

Extracellular Matrix Proteins ◽

Polyclonal Antiserum ◽

Glycolytic Enzymes ◽

Matrix Proteins ◽

Dependent Manner ◽

Human Respiratory Tract ◽

Concentration Dependent Manner ◽

Micro Organisms

In different, phylogenetically unrelated micro-organisms, glycolytic enzymes play a dual role. In the cytosol they are involved in metabolic reactions whereas the surface-localized fraction of the enzymes contributes to adhesion and virulence. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a typical member of this group of multifunctional proteins. In this study, we characterized the GAPDH of Mycoplasma pneumoniae, a common pathogen of the human respiratory mucosa. Full-length GAPDH of M. pneumoniae was successfully expressed and used to produce a polyclonal antiserum. By immunofluorescence, colony blot and ELISA experiments with different fractions of the M. pneumoniae proteins, GAPDH was demonstrated to be present in the cytosol and at even higher concentrations at the surface of mycoplasmas. Nevertheless, antibodies against recombinant GAPDH were not detected in sera of immunized animals or of patients with confirmed M. pneumoniae infection. Recombinant GAPDH bound to different human cell lines in a concentration-dependent manner, and binding was inhibited by specific anti-GAPDH serum. In contrast, this antiserum did not significantly influence the adherence of M. pneumoniae to HeLa cells. When different human extracellular matrix proteins were tested in Western blot assays, GAPDH bound to fibrinogen. The results showed that the GAPDH of M. pneumoniae is a member of the family of cytosol-localized glycolytic enzymes, which also occur at the surface of the bacterium, and mediates interactions with the extracellular matrix proteins of the human host. Thus, the surface-exposed fraction of GAPDH may be a factor that contributes to the successful colonization of the human respiratory tract by M. pneumoniae.

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Toxicity of Industrially Relevant Chlorinated Organic Solvents In Vitro

International Journal of Toxicology ◽

10.1177/1091581813482006 ◽

2013 ◽

Vol 32 (2) ◽

pp. 136-145 ◽

Cited By ~ 13

Author(s):

Catherine McDermott ◽

James J.A. Heffron

Keyword(s):

Cell Proliferation ◽

Organic Solvents ◽

Caspase 3 ◽

Free Calcium ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Chlorinated Organic ◽

Free Calcium Concentration ◽

Ros Formation

The cytotoxic effects of 4 industrially important chlorinated organic solvents, dichloromethane (DCM), 1,2-dichloroethane (DCE), trichloroethylene (TCE), and tetrachloroethylene (PERC) in vitro, were investigated. Jurkat T cells were exposed to the solvents individually for 72 hours and changes in reactive oxygen species (ROS) formation, cell proliferation, intracellular free calcium concentration ([Ca2+]), and caspase-3 activity were measured. There was a concentration-dependent increase in the ROS formation and intracellular free [Ca2+] following exposure to each of the solvents. This was accompanied by a decrease in the cell proliferation. Solvent potency decreased in the following order: PERC > TCE > DCM > DCE. Caspase-3 activity was increased in a concentration-dependent manner by TCE and PERC but was not significantly altered by DCM or DCE. n-Acetyl-l-cysteine pretreatment showed that changes in the intracellular free [Ca2+] and caspase-3 activity were independent of ROS formation. However, increased ROS formation did play a causal role in the decreased cell proliferation observed.

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Isoliquiritigenin inhibits the survival of diffuse large B-cell lymphoma cells by regulating Akt/mTOR signaling pathway

Tropical Journal of Pharmaceutical Research ◽

10.4314/tjpr.v19i8.8 ◽

2020 ◽

Vol 19 (8) ◽

pp. 1619-1623

Author(s):

Zhixiang Su ◽

Bin Yu ◽

Zhiping Deng ◽

Haifeng Sun

Keyword(s):

Cell Proliferation ◽

B Cell ◽

Signaling Pathway ◽

Cell Lymphoma ◽

B Cell Lymphoma ◽

Dependent Manner ◽

Mtor Signaling Pathway ◽

Concentration Dependent Manner ◽

Large B Cell Lymphoma ◽

Large B Cell

Purpose: To investigate the effect of isoliquiritigenin (ISL) on diffuse large B-cell lymphoma (DLBCL) cells and its underlying mechanism of action.Methods: The DLBCL cell line OCI-Ly19 was used in this study. Cell proliferation was measured by MTT assay. Apoptosis was evaluated using flow cytometry. Phosphorylation of Akt and mTOR was assessed using Western blotting.Results: DLBCL cell proliferation was suppressed by ISL in a concentration-dependent manner. The number of apoptotic cells increased following ISL treatment in a concentration-dependent manner (p < 0.05). ISL treatment also stopped the cell cycle at the G1 phase in a concentration-dependent manner. Western blot analysis indicated that there was no significant Akt and mTOR expression in cells treated with 10, 20, or 50 μM ISL (p < 0.05). However, Akt and mTOR phosphorylation was upregulated following treatment with 10, 20, or 50 μM ISL in a concentration-dependent manner (p < 0.05).Conclusion: The results demonstrate that ISL inhibits DLBCL cell proliferation and promotes cell apoptosis by blocking the cell cycle transition from the G1 to S phase, which is mediated by the inactivation of the Akt/mTOR signaling pathway. Keywords: Isoliquiritigenin, Cell survival, Diffuse large B-cell lymphoma, Akt/mTOR signaling pathway

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Adenosine A2A receptors promote adenosine-stimulated wound healing in bronchial epithelial cells

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00373.2005 ◽

2006 ◽

Vol 290 (5) ◽

pp. L849-L855 ◽

Cited By ~ 32

Author(s):

D. S. Allen-Gipson ◽

J. Wong ◽

J. R. Spurzem ◽

J. H. Sisson ◽

T. A. Wyatt

Keyword(s):

Wound Healing ◽

Epithelial Cells ◽

Adenosine Receptor ◽

Wound Repair ◽

Wound Closure ◽

Bronchial Epithelial Cells ◽

Dependent Manner ◽

Closure Rate ◽

Concentration Dependent Manner ◽

Bronchial Epithelial

Adenosine produces a wide variety of physiological effects through the activation of specific adenosine receptors (A1, A2A, A2B, A3). Adenosine, acting particularly at the A2A adenosine receptor (A2AAR), is a potent endogenous anti-inflammatory agent and sensor of inflammatory tissue damage. The complete healing of wounds is the final step in a highly regulated response to injury. Recent studies on epidermal wounds have identified the A2AAR as the main adenosine receptor responsible for altering the kinetics of wound closure. We hypothesized that A2AAR promotes wound healing in bronchial epithelial cells (BECs). To test this hypothesis, the human BEC line BEAS-2B and bovine BECs (BBECs) were used. Real-time RT-PCR of RNA from unstimulated BEAS-2B cells revealed transcriptional expression of A1, A2A, A2B and A3 receptors. Western blot analysis of lysates from BEAS-2B cells and BBECs detected a single band at 44.7 kDa in both the BECs, indicating the presence of A2AAR. In a wound healing model, we found that adenosine stimulated wound repair in cultured BBECs in a concentration-dependent manner, with an optimal closure rate observed between 4 and 6 h. Similarly, the A2AAR agonist 5′-( N-cyclopropyl)carboxamidoadenosine (CPCA) augmented wound closure, with a maximal closure rate occurring between 4 and 6 h. Inhibition of A2AAR with ZM-241385, a known A2AAR antagonist, impeded wound healing. In addition, ZM-241385 also attenuated adenosine-mediated wound repair. Kinase studies revealed that adenosine-stimulated airway repair activates PKA by ligating A2AAR. Collectively, the data suggest that the A2AAR is involved in BEC adenosine-stimulated wound healing and may prove useful in understanding purinergic-mediated actions on airway epithelial repair.

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Involvement of P1 Adhesin in Gliding Motility of Mycoplasma pneumoniae as Revealed by the Inhibitory Effects of Antibody under Optimized Gliding Conditions

Journal of Bacteriology ◽

10.1128/jb.187.5.1875-1877.2005 ◽

2005 ◽

Vol 187 (5) ◽

pp. 1875-1877 ◽

Cited By ~ 70

Author(s):

Shintaro Seto ◽

Tsuyoshi Kenri ◽

Tetsuo Tomiyama ◽

Makoto Miyata

Keyword(s):

Monoclonal Antibody ◽

Mycoplasma Pneumoniae ◽

Conformational Changes ◽

Gliding Motility ◽

Dependent Manner ◽

Inhibitory Effects ◽

Slight Effect ◽

Concentration Dependent Manner ◽

Gliding Mechanism ◽

Over Time

ABSTRACT To examine the participation of P1 adhesin in gliding of Mycoplasma pneumoniae, we examined the effects of an anti-P1 monoclonal antibody on individual gliding mycoplasmas. The antibody reduced the gliding speed and removed the gliding cells from the glass over time in a concentration-dependent manner but had only a slight effect on nongliding cells, suggesting that the conformational changes of P1 adhesin and its displacement are involved in the gliding mechanism.

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In Vitro Evidence for Effects of Magnesium Supplementation on Quinolone-treated Horse and Dog Chondrocytes

Veterinary Pathology ◽

10.1354/vp.38-2-143 ◽

2001 ◽

Vol 38 (2) ◽

pp. 143-148 ◽

Cited By ~ 22

Author(s):

M. Egerbacher ◽

B. Wolfesberger ◽

C. Gabler

Keyword(s):

Cell Proliferation ◽

Magnesium Deficiency ◽

Great Part ◽

Control Group ◽

Dependent Manner ◽

Free Medium ◽

Culture Dish ◽

Magnesium Supplementation ◽

Concentration Dependent Manner

Quinolones and magnesium deficiency cause similar lesions in joint cartilage of young animals. Chondrocytes cultivated in the presence of quinolones and in Mg-free medium show severe alterations in cytoskeleton and decreased ability to adhere to the culture dish. We investigated whether Mg2+ supplementation can prevent quinolone-mediated effects on chondrocytes in vitro. Chondrocytes cultivated in Dulbecco's modified Eagle's medium/HAM's F-12 medium were treated with ciprofloxacin (80 and 160 μg/ml) and enrofloxacin (100 and 150 μg/ml). Mg2+ was added at a concentration of 0.0612 mg/ml (MgCl) and 0.0488 mg/ml (MgSO4) or a triple dose. In addition, cells were cultivated in Mg-free medium and accordingly treated with Mg2+ supplementation. After 5 days in culture, the number of adherent cells per milliliter was determined. The number of chondrocytes in quinolone-treated groups decreased to 12-36% that of the control group within the culture period. With Mg2+ supplementation, the number of attached cells increased to 40-70% that of control cells. The threefold dose of Mg2+ led to better results than did the single dose. Cell proliferation tested by immunohistochemical staining with Ki67 (clone MIB5) decreased from 70% in control groups to 55%, 48%, and 30% in enrofloxacintreated groups in a concentration dependent manner (50, 100, and 150 μg/ml). Addition of Mg2+ did not increase the rate of cell proliferation. These results suggest that a great part of quinolone-induced damage is due to magnesium complex formation, as Mg2+ supplementation is able to reduce the effects in vitro. However, quinolone effects on cell proliferation seem to be an independent process that is not influenced by magnesium supplementation.

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