scholarly journals Application of Humanized Zebrafish Model in the Suppression of SARS-CoV-2 Spike Protein Induced Pathology by Tri-Herbal Medicine Coronil via Cytokine Modulation

Molecules ◽  
2020 ◽  
Vol 25 (21) ◽  
pp. 5091
Author(s):  
Acharya Balkrishna ◽  
Siva Kumar Solleti ◽  
Sudeep Verma ◽  
Anurag Varshney
Keyword(s):  
Herbal Medicine ◽  
High Performance ◽  
Normal Skin ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Spike Protein ◽  
Protective Effects ◽  
Cell Degeneration ◽  
Zebrafish Model ◽  
Tnf Α

Zebrafish has been a reliable model system for studying human viral pathologies. SARS-CoV-2 viral infection has become a global chaos, affecting millions of people. There is an urgent need to contain the pandemic and develop reliable therapies. We report the use of a humanized zebrafish model, xeno-transplanted with human lung epithelial cells, A549, for studying the protective effects of a tri-herbal medicine Coronil. At human relevant doses of 12 and 58 µg/kg, Coronil inhibited SARS-CoV-2 spike protein, induced humanized zebrafish mortality, and rescued from behavioral fever. Morphological and cellular abnormalities along with granulocyte and macrophage accumulation in the swim bladder were restored to normal. Skin hemorrhage, renal cell degeneration, and necrosis were also significantly attenuated by Coronil treatment. Ultra-high-performance liquid chromatography (UHPLC) analysis identified ursolic acid, betulinic acid, withanone, withaferine A, withanoside IV–V, cordifolioside A, magnoflorine, rosmarinic acid, and palmatine as phyto-metabolites present in Coronil. In A549 cells, Coronil attenuated the IL-1β induced IL-6 and TNF-α cytokine secretions, and decreased TNF-α induced NF-κB/AP-1 transcriptional activity. Taken together, we show the disease modifying immunomodulatory properties of Coronil, at human equivalent doses, in rescuing the pathological features induced by the SARS-CoV-2 spike protein, suggesting its potential use in SARS-CoV-2 infectivity.

Dual mechanism forMycobacterium tuberculosiscytotoxicity on lung epithelial cells

2012 ◽  
Vol 58 (7) ◽  
pp. 909-916 ◽  
Author(s):  
Jorge Castro-Garza ◽  
W. Edward Swords ◽  
Russell K. Karls ◽  
Frederick D. Quinn
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Neutralizing Antibodies ◽  
Cytotoxic Effect ◽  
Alveolar Epithelial Cell ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Necrosis Factor Alpha ◽  
Tnf Α ◽  
Lung Epithelial

Mycobacterium tuberculosis strains CDC1551 and Erdman were used to assess cytotoxicity in infected A549 human alveolar epithelial cell monolayers. Strain CDC1551 was found to induce qualitatively greater disruption of A549 monolayers than was strain Erdman, although total intracellular and cell-associated bacterial growth rates over the course of the infections were not significantly different. Cell-free culture supernatants from human monocytic cells infected with either of the 2 M. tuberculosis strains produced a cytotoxic effect on A549 cells, correlating with the amount of tumor necrosis factor alpha (TNF-α) released by the infected monocytes. The addition of TNF-α-neutralizing antibodies to the supernatants from infected monocyte cultures did prevent the induction of a cytotoxic effect on A549 cells overlaid with this mixture but did not prevent the death of epithelial cells when added prior to infection with M. tuberculosis bacilli. Thus, these data agree with previous observations that lung epithelial cells infected with M. tuberculosis bacilli are rapidly killed in vitro. In addition, the data indicate that some of the observed epithelial cell killing may be collateral damage; the result of TNF-α released from M. tuberculosis-infected monocytes.

scholarly journals Induction of Proinflammatory Cytokines in Human Lung Epithelial Cells during Chlamydia pneumoniae Infection

2003 ◽  
Vol 71 (2) ◽  
pp. 614-620 ◽  
Author(s):  
Jun Yang ◽  
W. Craig Hooper ◽  
Donald J. Phillips ◽  
Maria L. Tondella ◽  
Deborah F. Talkington
Keyword(s):  
Epithelial Cells ◽  
Chlamydia Pneumoniae ◽  
Human Lung ◽  
A549 Cells ◽  
Cytokine Response ◽  
Lung Epithelial Cells ◽  
Cytokine Gene Expression ◽  
Cytokine Release ◽  
Tnf Α ◽  
Respiratory Epithelial

ABSTRACT Chlamydia pneumoniae is an obligate intracellular human pathogen that causes acute respiratory diseases such as pneumonia and bronchitis. Previous studies have established that C. pneumoniae can induce cytokines in mouse and/or human cells, but little information is available on the cytokine response of respiratory epithelial cells, a first line of infection. In this study, heparin treatment of C. pneumoniae significantly reduced its ability to induce interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-α) mRNA in human lung carcinoma cells, indicating that cytadherence is an important early stimulus for induction of proinflammatory mediators. Although the IL-8, gamma interferon, and TNF-α message was consistently induced by infection of A549 cells not treated with heparin, only an elevation of IL-8 protein was detected in A549 supernatants. A549 IL-β and IL-6 mRNA and supernatant protein profiles were not significantly changed by infection. Heat or UV inactivation of C. pneumoniae only partially reduced the cytokine response, and inhibition of C. pneumoniae protein or DNA synthesis did not affect its ability to induce cytokine gene expression. To prevent stress-induced cytokine release by the A549 cells, centrifugation was not utilized for infection experiments. These experiments establish the importance of cytadherence in cytokine release by cells of respiratory epithelial origin and suggest that further work in the area of cytokine mediators is warranted to gain valuable pathogenic and therapeutic insights.

Contribution of IL-1β and TNF-α to the initiation of the peripheral lung response to atmospheric particulates (PM10)

2004 ◽  
Vol 287 (1) ◽  
pp. L176-L183 ◽  
Author(s):  
Hiroshi Ishii ◽  
Takeshi Fujii ◽  
James C. Hogg ◽  
Shizu Hayashi ◽  
Hiroshi Mukae ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Vegf Mrna ◽  
Atmospheric Particulates ◽  
Proinflammatory Mediators ◽  
Acute Response ◽  
Chemotactic Protein ◽  
Tnf Α ◽  
Macrophage Colony

Alveolar macrophages (AM) play a key role in clearing atmospheric particulates from the lung surface and stimulating epithelial cells to produce proinflammatory mediators. The present study examines the role of “acute response” cytokines TNF-α and IL-1β released by AM exposed to ambient particulate matter with a diameter of <10 μm (PM10) in amplifying the proinflammatory mediator expression by A549 cells and human bronchial epithelial cells (HBEC). The results showed that supernatants from human AM incubated 24 h with PM10(100 μg/ml) contained more TNF-α, IL-1β, granulocyte-macrophage colony stimulating factor, IL-6, and IL-8 than nonexposed AM supernatants. The 3-h treatment of A549 cells with PM10-exposed AM supernatants increased TNF-α, IL-1β, IL-8, regulated on activation normal T-cells expressed and secreted (RANTES), and leukemia inhibitory factor mRNA compared with the treatment with nonexposed AM supernatants and, compared with untreated A549 cells, additionally increased ICAM-1 and monocyte chemotactic protein-1 mRNA. Preincubating PM10-exposed AM supernatants with anti-IL-1β antibodies reduced all the above mediators as well as VEGF mRNA expression ( P < 0.05), while anti-TNF-α antibodies were less effective ( P > 0.05), and the combination of the two antibodies most effective. When HBEC were treated similarly, anti-TNF-α antibodies had the greatest effect. In A549 cells PM10-exposed AM supernatants increased NF-κB, activator protein (AP)-1 and specificity protein 1 binding, while anti-TNF-α and anti-IL-1β antibodies reduced NF-κB and AP-1 binding. We conclude that AM-derived TNF-α and IL-1β provide a major stimulus for the production of proinflammatory mediators by lung epithelial cells and that their relative importance may depend on the type of epithelial cell target.

scholarly journals Protective effects of wogonin on lipopolysaccharide-induced inflammation and apoptosis of lung epithelial cells and its possible mechanisms

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Jinlin Ge ◽  
Huanhuan Yang ◽  
Yufeng Zeng ◽  
Yunjie Liu
Keyword(s):  
Oxidative Stress ◽  
Western Blot ◽  
A549 Cells ◽  
High Mobility ◽  
Lung Epithelial Cells ◽  
Sirtuin 1 ◽  
Protective Effects ◽  
Sirt1 Inhibitor ◽  
Regulatory Effects

Abstract Background Wogonin (5, 7-dihydroxy-8-methoxyflavone) is a natural di-hydroxyl flavonoid extracted from the root of Scutellaria baicalensis Georgi. This paper was intended to investigate the mechanism of action of wogonin in alleviating the inflammation and apoptosis in acute lung injury (ALI). Materials and methods Lipopolysaccharide (LPS) was used to establish the in vitro model of ALI. After wogonin treatment, the cell viability and apoptosis of LPS-induced A549 cells were, respectively, measured by CCK-8, TUNEL assays and acridine orange/ethidium bromide dual staining, while the contents of inflammatory cytokines and oxidative stress markers were estimated by RT-qPCR, ELISA assay, western blot analysis and commercial kits. Western blot was also conducted to assess the expression of proteins involved. Subsequently, the effect of wogonin on the sirtuin 1 (SIRT1)-mediated high-mobility group box 1 protein (HMGB1) deacetylation was investigated. SIRT1 inhibitor EX527 was used to evaluate the regulatory effects of wogonin on SIRT1-mediated HMGB1 deacetylation in A549 cells under LPS stimulation. Results LPS induced inflammation, oxidative stress and apoptosis of A549 cells, which was abolished by wogonin. It was also found that wogonin promoted the HMGB1 deacetylation, accompanied by upregulated SIRT1 expression. However, SIRT1 inhibitor EX527 partially reversed the protective effects of wogonin on the inflammation and apoptosis of LPS-induced A549 cells. Conclusion Wogonin alleviated the inflammation and apoptosis in LPS-induced A549 cells by SIRT1-mediated HMGB1 deacetylation, which might represent the identification of a novel mechanism by which wogonin exerts protective effects on ALI and provide ideas for the application of wogonin to ALI treatment.

scholarly journals Quercetin Prevents LPS-Induced Oxidative Stress and Inflammation by Modulating NOX2/ROS/NF-kB in Lung Epithelial Cells

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6949
Author(s):  
 Ok-Joo Sul ◽  
Seung Won Ra
Keyword(s):  
Oxidative Stress ◽  
Nuclear Translocation ◽  
A549 Cells ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Lung Epithelial Cells ◽  
Tnf Α ◽  
Lung Epithelial ◽  
Cytokine Tumor Necrosis Factor ◽  
Inflammatory Processes ◽  
Mrna And Protein Expression

Oxidative stress caused by the production of reactive oxygen species (ROS) plays a major role in inflammatory processes. We hypothesized that modulation of ROS via quercetin may protect against oxidative stress and inflammation. Thus, this study aimed to investigate the effects of quercetin on oxidative stress and inflammation in lung epithelial A549 cells. The lipopolysaccharide (LPS)-induced elevation of intracellular ROS levels was reduced after quercetin treatment, which also almost completely abolished the mRNA and protein expression of nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2) induced by LPS stimulation. In addition, quercetin suppressed the nuclear translocation of nuclear factor kappa B (NF-κB) and reduced levels of inflammatory cytokine tumor necrosis factor (TNF)-α, interleukin (IL)-1, and IL-6, which had increased significantly after LPS exposure. Our data demonstrated that quercetin decreased ROS-induced oxidative stress and inflammation by suppressing NOX2 production.

The effects of PM10 particles and oxidative stress on macrophages and lung epithelial cells: modulating effects of calcium-signaling antagonists

2007 ◽  
Vol 292 (6) ◽  
pp. L1444-L1451 ◽  
Author(s):  
David M. Brown ◽  
Laura Hutchison ◽  
Kenneth Donaldson ◽  
Vicki Stone
Keyword(s):  
Oxidative Stress ◽  
Calcium Antagonist ◽  
Epithelial Cells ◽  
Calcium Signaling ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Tnf Α ◽  
Lung Epithelial

We have previously examined the ability of air pollution particles (PM10) to promote release of the proinflammatory cytokine tumor necrosis factor-α (TNF-α) from human peripheral blood mononuclear cells and demonstrated a role for calcium as a signaling molecule in this process. We have now studied the ability of oxidative stress induced by a synthetic oxidant tert-butyl hydroperoxide (tBHP) to induce TNF-α production via calcium signaling in the mouse macrophage cell line (J774). The oxidant tBHP significantly increased intracellular calcium and the release of TNF-α in J774 cells, an effect that was reduced to control levels by inhibition of calcium signaling with verapamil, BAPTA-AM, and W-7. This study also investigated interactions between PM10-treated macrophages and epithelial cells by using conditioned medium (CM) from PM10-treated mononuclear cells to stimulate the release of the neutrophil chemoattractant chemokine IL-8 from A549 lung epithelial cells. TNF-α protein release was demonstrated in human mononuclear cells after PM10 treatment, an effect that was inhibited by calcium antagonists. Treatment of A549 cells with monocyte/PM10 CM produced increased IL-8 release that was reduced with CM from monocyte/PM10/calcium antagonist treatments. The expression of ICAM-1 was increased after incubation with CM from monocyte/PM10 treatment, and this increase was prevented by treatment with CM from monocyte/PM10/calcium antagonist. These data demonstrate a link between oxidative stress, calcium, and inflammatory mediator production in macrophages and lung epithelial cells.

TNF-α induces MUC1 gene transcription in lung epithelial cells: its signaling pathway and biological implication

2007 ◽  
Vol 293 (3) ◽  
pp. L693-L701 ◽  
Author(s):  
Takeshi Koga ◽  
Ippei Kuwahara ◽  
Erik P. Lillehoj ◽  
Wenju Lu ◽  
Takeshi Miyata ◽  
...  
Keyword(s):  
Gene Transcription ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Luciferase Reporter ◽  
Promoter Activation ◽  
Alveolar Cells ◽  
Intact Cells ◽  
Protein Levels ◽  
Muc1 Gene ◽  
Tnf Α

The current study was conducted to elucidate the mechanism through which TNF-α stimulates expression of MUC1, a membrane-tethered mucin. A549 human lung alveolar cells treated with TNF-α exhibited significantly higher MUC1 protein levels in detergent lysates compared with cells treated with vehicle alone. Increased MUC1 protein levels were correlated with significantly higher levels of MUC1 mRNA in TNF-α-treated cells compared with controls. However, TNF-α did not alter MUC1 transcript stability, implying increased de novo transcription induced by the cytokine. TNF-α increased MUC1 gene promoter activity in A549 cells transfected with a promoter-luciferase reporter plasmid. Both U0126, an inhibitor of MEK1/2, and dominant negative ERK1 prevented TNF-α-induced MUC1 promoter activation, and anti-TNFR1 antibody blocked TNF-α-stimulated ERK1/2 activation. MUC1 promoter activation by TNF-α also was blocked by mithramycin A, an inhibitor of Sp1, as well as either deletion or mutation of a putative Sp1 binding site in the MUC1 promoter located between nucleotides −99 and −90. TNF-α-stimulated binding of Sp1 to the MUC1 promoter in intact cells was demonstrated by chromatin immunoprecipitation assay. We conclude that TNF-α induces MUC1 gene transcription through a TNFR1 → MEK1/2 → ERK1 → Sp1 pathway.

scholarly journals Giloy Ghanvati (Tinospora cordifolia (Willd.) Hook. f. and Thomson) Reversed SARS-CoV-2 Viral Spike-Protein Induced Disease Phenotype in the Xenotransplant Model of Humanized Zebrafish

2021 ◽  
Vol 12 ◽  
Author(s):  
Acharya Balkrishna ◽  
Lakshmipathi Khandrika ◽  
Anurag Varshney
Keyword(s):  
Treatment Option ◽  
Symptomatic Relief ◽  
Viral Disease ◽  
Lung Epithelial Cells ◽  
Spike Protein ◽  
Tinospora Cordifolia ◽  
Swim Bladder ◽  
Disease Phenotype ◽  
Zebrafish Model ◽  
Holistic Treatment

The current Severe Acute Respiratory Syndrome disease caused by Coronavirus-2 (SARS-CoV-2) has been a serious strain on the healthcare infrastructure mainly due to the lack of a reliable treatment option. Alternate therapies aimed at symptomatic relief are currently prescribed along with artificial ventilation to relieve distress. Traditional medicine in the form of Ayurveda has been used since ancient times as a holistic treatment option rather than targeted therapy. The practice of Ayurveda has several potent herbal alternatives for chronic cough, inflammation, and respiratory distress which are often seen in the SARS-CoV-2 infection. In this study we have used the aqueous extracts of Tinospora cordifolia (willd.) Hook. f. and Thomson in the form of Giloy Ghanvati, as a means of treatment to the SARS-CoV-2 spike-protein induced disease phenotype in a humanized zebrafish model. The introduction of spike-protein in the swim bladder transplanted with human lung epithelial cells (A549), caused an infiltration of pro-inflammatory immune cells such as granulocytes and macrophages into the swim bladder. There was also an increased systemic damage as exemplified by renal tissue damage and increased behavioral fever in the disease induction group. These features were reversed in the treatment group, fed with three different dosages of Giloy Ghanvati. The resultant changes in the disease phenotype were comparable to the group that were given the reference compound, Dexamethasone. These findings correlated well with various phyto-compounds detected in the Giloy Ghanvati and their reported roles in the viral disease phenotype amelioration.

Pneumocystis cariniiinduces ICAM-1 expression in lung epithelial cells through a TNF-α-mediated mechanism

1997 ◽  
Vol 273 (6) ◽  
pp. L1103-L1111 ◽  
Author(s):  
Mariette L. Yu ◽  
Andrew H. Limper
Keyword(s):  
Epithelial Cells ◽  
Pneumocystis Carinii ◽  
A549 Cells ◽  
Lung Epithelial Cells ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Respiratory Impairment ◽  
Type Ii Pneumocytes ◽  
Tnf Α ◽  
Factor Α

Inflammatory cell recruitment contributes to respiratory impairment during Pneumocystis carinii pneumonia. We evaluated expression of intercellular adhesion molecule-1 (ICAM-1), a key participant in leukocyte accumulation, in rats with P. carinii pneumonia. Immunostaining for ICAM-1 was most marked on bronchiolar epithelium but was also evident on type II pneumocytes, endothelium, and macrophages. Lung from normal and dexamethasone-treated uninfected animals exhibited markedly less ICAM-1. We hypothesized that P. carinii promoted ICAM-1 expression in epithelium through tumor necrosis factor-α (TNF-α) release from macrophages or that P. carinii directly stimulated ICAM-1 expression. Alveolar macrophages were incubated with P. carinii, and the medium was added to A549 epithelial cells. Treatment of macrophages with P. carinii enhanced A549 ICAM-1, which was inhibited with antibody to TNF-α. To determine whether P. carinii alone also stimulated ICAM-1, A549 cells were cultured with P. carinii, also augmenting ICAM-1. Of note, A549 ICAM-1 expression from P. carinii alone was less than with P. carinii-exposed macrophages. Thus ICAM-1 is enhanced in lung epithelium during P. carinii infection, in part, through TNF-α-mediated mechanisms.

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